CN108048338A - One Accharomyces cerevisiae and its application in aquaculture - Google Patents
One Accharomyces cerevisiae and its application in aquaculture Download PDFInfo
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- CN108048338A CN108048338A CN201711493877.9A CN201711493877A CN108048338A CN 108048338 A CN108048338 A CN 108048338A CN 201711493877 A CN201711493877 A CN 201711493877A CN 108048338 A CN108048338 A CN 108048338A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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Abstract
Application the invention discloses an Accharomyces cerevisiae and its in aquaculture, belongs to microbial technology field.The saccharomyces cerevisiae deposit number of the present invention is that deposit number is CCTCC NO：The saccharomyces cerevisiae F2 of M2017792, the bacterial strain being capable of degradation of ammonia nitrogen, can in breeding water body rapid conversion ammonia nitrogen into protein, generate a large amount of thalline provides bait for aquatic animal simultaneously, maintain aquatic livestock intestinal microecology balance, aquatic livestock survival rate and yield are improved, there is industrialized developing and the potentiality promoted and applied.
Description
Technical field
The present invention relates to microbial technology fields, and in particular to an Accharomyces cerevisiae and its application in aquaculture.
Background technology
With industrial pollution discharge, livestock and poultry breeding industry sewage discharge, sanitary sewage discharge, excessively feed feeding in aquaculture
Material behavior etc., the exceeded thing that aquatile is caused to be poisoned to death of ammonia nitrogen frequently occurs in breeding water body, and pole is brought to raiser
Big economic loss.
Total ammonia nitrogen in breeding water body is generally non-ionic ammonia (NH in two forms3) and ammonium ion (NH4 +) exist, in pH
When value is less than 7, the ammonia in water is nearly all with NH4 +Form exist, pH be more than 11 when, then nearly all with NH3Form deposit
, temperature rise, NH3Ratio increase.Ammonia nitrogen is primarily referred to as the harm of aquatile the harm of non-ionic ammonia, nonionic
After ammonia enters in aquatile body, enzymatic hydrolysis reaction and membrane stability generation are significantly affected, expiratory dyspnea is shown, does not take the photograph
Phenomena such as food, resistance decline, faint from fear, stupor, even results in the large quantities of death of aquatile.In addition, be enriched in vivo
Ammonia nitrogen in high density can be converted into after nitrite and generate harm to organism, and nitrite is strong oxidizer, can not only be made
Organism toxicosis, it also has carcinogenesis.According to《Water quality standard for fishery》, in aquaculture production, the concentration of ammonia should be controlled
In below 0.02mg/L.
The content of the invention
Application it is an object of the invention to provide an Accharomyces cerevisiae and its in aquaculture, the bacterial strain can cultivate
Rapid conversion ammonia nitrogen is into protein in water body, while generates a large amount of thalline and provide bait for aquatic animal, improves aquatic livestock and deposits
Motility rate and yield have industrialized developing and the potentiality promoted and applied.
The purpose of the present invention is achieved through the following technical solutions：
One Accharomyces cerevisiae is saccharomyces cerevisiae (Saccharomyces cerevisiae) F2, and the bacterial strain is in 2017
December 15 was preserved in China typical culture collection center (address：Chinese Wuhan Wuhan Universitys), Classification And Nomenclature is：
Saccharomyces cerevisiae F2 S. cervisiae F2, deposit number are：CCTCC NO：M2017792.
The Yeast protoplase and physiological and biochemical property：The bacterium colony that the bacterial strain breeds formation on YPD solid mediums is circular, greatly
And moisten, it is creamy white, surface elevation is smooth non-wrinkled, edge clear, and growth is rapid；Microscopy shows that cell individual is larger,
In subsphaeroidal, polygon budding can generate ascospore.Glucose fermentation, galactolipin, maltose, sucrose, raffinose fermentation 1/
3, do not assimilate nitrate, not decomposing urea, produce ester, resistance to 10% ethyl alcohol.
The saccharomyces cerevisiae can be used in aquaculture, and ammonia nitrogen in breeding water body can be quickly reduced using saccharomyces cerevisiae F2
Concentration safeguards water body microecological balance.
Additive amount of the saccharomyces cerevisiae in aquaculture process adds 200-400g for every meter of depth of water per acre, described
Saccharomyces cerevisiae living bacteria count be hundred million CFU/g of 100-150.
Compared with prior art, the present invention has the following advantages：The saccharomyces cerevisiae F2 of the present invention can be reduced in breeding water body
Ammonia-nitrogen content can maintain aquatic livestock intestinal microecology to balance, and promote growth.
Description of the drawings
Fig. 1 be in embodiment 1 different isolated strains to the removal effect figure of ammonia nitrogen.
Fig. 2 be in embodiment 2 different carbon source to the influence result figure of saccharomyces cerevisiae F2 degradation of ammonia nitrogen.
Fig. 3 be in embodiment 2 saccharomyces cerevisiae F2 to the graph of ammonia nitrogen degradation.
Specific embodiment
The present invention is described in further details with reference to specific embodiment.Embodiment using technical solution of the present invention as
Under the premise of implemented, give detailed embodiment and specific operating process, but protection scope of the present invention is not limited to
Following embodiments.Method therefor is conventional method unless otherwise instructed in following embodiments.
The screening and identification of 1 saccharomyces cerevisiae of embodiment
(1) bacterial strain primary dcreening operation
The water sampling from Wuhan Tian occasion bio tech ltd Pelteobagrus fulvidraco aquaculture pond is transferred to containing 50 μ g/mL ammonia benzyls
30 DEG C of aerobic enrichment cultures are muddy in the YPD fluid nutrient mediums of penicillin, apply tablet after appropriate dilution, chosen according to colonial morphology
Take products of typical yeast bacterium single bacterium colony (white, size 1-2mm, moistening protuberance have ester fragrance).
YPD fluid nutrient mediums：Glucose 20g, tryptone 20g, dusty yeast 10g, water 1000mL adjust 7.0,115 DEG C of pH
Sterilize 20min.
YPD solid mediums：Glucose 20g, tryptone 20g, dusty yeast 10g, agar 18g, water 1000mL adjust pH
7.0,115 DEG C of sterilizing 20min.
(2) bacterial strain secondary screening
The single bacterium colony that primary dcreening operation is obtained is inoculated into the sterilizing breeding water body (121 DEG C of 30min) that ammonia nitrogen concentration is 5.3mg/L
In, 25 DEG C of cultures measure its NH afterwards for 24 hours4 +Concentration, the result is shown in Figure 1, screening obtain the excellent saccharomycete F2 of one plant of character.
(3) identify
Reference《Fungal identification handbook》Individual morphology observation, biochemical characteristic experiment, growth spy are carried out to the bacterial strain screened
Property experiment and DNA identifications, determine that final screening obtains is an Accharomyces cerevisiae.
F2 bacterial strains bacterium colony and thalli morphology：The bacterium colony that the bacterial strain breeds formation on YPD solid mediums is circular, big and wet
Profit, is creamy white, and surface elevation is smooth non-wrinkled, edge clear, and growth is rapid；Microscopy shows that cell individual is larger, near
Spherical shape, polygon budding can generate ascospore.
Physiological and biochemical property：Glucose fermentation, galactolipin, maltose, sucrose, raffinose fermentation 1/3, do not assimilate nitric acid
Salt, decomposing urea, does not produce ester, resistance to 10% ethyl alcohol.
The genomic DNA of F2 bacterial strains is extracted, PCR amplification 26SrDNA D1/D2 areas carry out molecular level identification, will
26SrDNA D1/D2 areas extension increasing sequence (sequencing result is shown in SEQ ID NO.1) and the saccharomyces cerevisiae reported on GeneBanK
26SrDNA D1/D2 region sequences are compared, and homology reaches 100%, further confirm the bacterium as saccharomyces cerevisiae.Wherein,
PCR amplification sense primer：5`-GCATATCAATAAGCGGAGGAAAAC-3`, PCR amplification anti-sense primer： 5`-
GGTCCGTGTTTCAAGACGC-3`。
F2 bacterial strains are preserved in China typical culture collection center (address on December 15th, 2017：Chinese Wuhan
Wuhan University), Classification And Nomenclature is：Saccharomyces cerevisiae F2 S. cervisiae F2, deposit number are：
CCTCC NO： M2017792.
2 saccharomyces cerevisiae F2 of embodiment removes ammonia nitrogen experiment in breeding water body
(1) by saccharomyces cerevisiae F2 single bacterium colonies be inoculated into 50mL YPD fluid nutrient mediums 30 DEG C, 200rpm culture for 24 hours, from
The heart removes culture medium and is resuspended with 50mL sterile waters.Absorption 1.0mL resuspended bacterium solutions are seeded to 100mL and contain the cultivation of 45mg/L ammonia nitrogens
In water body, the glucose of addition final concentration 2g/L, sucrose, ethyl alcohol, sodium acetate, sodium citrate do not add carbon source as carbon source respectively
As a control group, every group of 3 repetitions, 25 DEG C of cultures measure ammonia nitrogen concentration for 24 hours, and the result is shown in Fig. 2.As shown in Figure 2, glucose and
Sucrose is the good sources of carbon that bacterial strain F2 carries out ammonia nitrogen degradation, and ammonia nitrogen degradation rate is up to more than 99% for 24 hours.
(2) by saccharomyces cerevisiae F2 single bacterium colonies be inoculated into 50mL YPD fluid nutrient mediums 30 DEG C, 200rpm culture for 24 hours, from
The heart removes culture medium and is resuspended with 50mL sterile waters.Absorption 0.5,1.0,1.5mL resuspended bacterium solutions are seeded to 100mL and contain respectively
In 10.4mg/L ammonia nitrogen breeding water bodies, addition final concentration 100mg/L glucose is not inoculated with as a control group, every group as carbon source
3 repetitions, at 25 DEG C culture measure ammonia nitrogen concentration every 1h, the result is shown in Fig. 3.From the figure 3, it may be seen that when glucose is as carbon source
Bacterial strain F2 has fast degradation ability to ammonia nitrogen in breeding water body, and when inoculum concentration is 1.5mL, degradation rate is up to 0.46mg/
L·h。
The expansion culture of 3 saccharomyces cerevisiae F2 of embodiment
(1) saccharomyces cerevisiae F2 single bacterium colonies are inoculated into 50mL YPD fluid nutrient mediums, 30 DEG C, 200rpm culture make for 24 hours
For primary seed solution.
(2) by primary seed solution transfer 2mL to 200mL YPD fluid nutrient mediums in, 30 DEG C, 200rpm culture 12h conducts
Secondary seed solution.
(3) 50L fermentation tanks expand culture
By 10L initial mediums (100g glucose, 50g ammonium sulfate, 10g potassium dihydrogen phosphates, 10L water, adjust pH to 6.0,
115 DEG C of sterilizing 20min) it is added in 50L fermentation tanks, it is inoculated with above-mentioned secondary seed solution and ferments.Using glucose as carbon source,
Ammonium sulfate supplements C, N, P, fermentation whole process adds glucose as phosphorus source as nitrogen source, potassium dihydrogen phosphate in a manner that stream adds
(0.225-0.4kg/L), add ammonium sulfate (30-32.5g/L), add potassium dihydrogen phosphate (5-6g/L), to send out per liter per hour
The speed for adding in 18mL in zymotic fluid is added into zymotic fluid, altogether stream plus 20L.In fermentation process pH5.0- is adjusted with sodium carbonate
6.0, fermentation process glucose content 1-10g/L is maintained, 37 DEG C of whole process of fermenting maintenance temperature and rotating speed 500rpm, aerlbic culture,
It is appropriate to adjust throughput to maintain dissolved oxygen >=5%.
Fermentation ends after 48h take and are counted under zymotic fluid blood counting chamber microscope, and bacteria concentration is 3.2 × 109Cfu/mL,
Zymotic fluid is collected after putting tank, precipitation is collected after low-speed centrifugal (5000rpm/min) 5min, after washing is resuspended in a small amount of sterile water,
Thalline were collected by centrifugation again (i.e. yeast paste), calculates to obtain thalline weight in wet base 180g/L, is 9,600,000,000 CFU per g yeast pastes living bacteria count.
Using fluidized bed drying, 75 DEG C of inlet air temperature, 50 DEG C of leaving air temp are controlled, the yeast dry mycelium living bacteria count of acquisition reaches
14000000000 or so CFU/g.
Applications of the 4 saccharomyces cerevisiae F2 of embodiment in Pelteobagrus fulvidraco cultivates
Contrast tests, examination are carried out in one farm of Foshan city Nanhai District Xi Qiao towns, four culture ponds of selection within 2016
Test the pool and equal two of the pool of control.Pond size is 8 mu, depth of water 2m of water surface area, and intensive culture Pelteobagrus fulvidraco in pond was put in 2 months
Seedling, cultivation density 40,000 tail per acre, stocking size are 20-30 tails/jin, pond water colour strong green, transparency 25cm.It is tested
When, two of which pond using saccharomyces cerevisiae F2 microbial inoculums (additive amount per acre every meter of depth of water 200g), other two pond as pair
According to not putting into this microbial inoculum.At 9 points in the pond morning is tested using this microbial inoculum, the 2nd day at 9 points in the morning was detected, and the results are shown in Table 1.Examination
The transparency for testing pond significantly improves, and ammonia nitrogen, nitrite concentration are decreased obviously, and dissolved oxygen content also substantially rises.And compare pond
Pool transparency, ammonia nitrogen, dissolved oxygen, nitrite are all without significant change.Pond waters pH value belongs to normal value between 8.2-8.6.
Saccharomyces cerevisiae F2 microbial inoculums can keep water body to be in preferable state 7 days using 1 time, and it is big that seedling amount is put in the cultivation of Pearl River Delta area Pelteobagrus fulvidraco,
It is more to feed feed, pond Assessment of Organic Pollution is serious, and ammonia nitrogen and nitrite are all easier to raise, and use saccharomyces cerevisiae F2 microbial inoculums
The preferable state of water quality is kept to belong to the long period in 7 days.Final mid-June sells fish crop：Persistently using saccharomyces cerevisiae F2 microbial inoculums
Experiment pond sells fish specification and be averaged 4.2 liang/tail, and per mu yield yield is close to 4500 jin, and the control of unused saccharomyces cerevisiae F2 microbial inoculums
Pond sells fish specification and is averaged 3.3 liang/tail, 3000 jin or so of per mu yield yield.Experiment proves saccharomyces cerevisiae F2 in Pelteobagrus fulvidraco cultivates,
Survival rate can not only be improved, moreover it is possible to reduce the Different Nitrogen Concentrations such as water body ammonia nitrogen, nitrite, and yellow catfish growing institute can be converted into
Bait is needed, improves cultured output.
Table 1 tests the pool with compareing pool water quality index
Sequence table
<110>Tian Chen bio tech ltd of Wuhan City
<120>One Accharomyces cerevisiae and its application in aquaculture
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 493
<212> DNA
<213> Saccharomyces cerevisiae
<400> 1
gcatatcaat aagcggagga aaactgaaac tgcgcatgag attgatgacg gcgtgtcggt 60
gaaaaactat ggtagagatg gactgtacac tttatatatt catattgcgg atccaacttc 120
tatgtttccg gaaagcacta atgttgatat tgagggtata agtacagata tcctaaatgt 180
tgccttgaaa agatcattta ccacatattt accagatacg gttgttccta tgttacctca 240
atctatttgt cacttatcag atttggggaa acaaggacaa aggacgaaaa ctatatcctt 300
ctctgttgat gttaaaatca cttctaaatg cagtggaaaa tcaattgaaa ttatgttatg 360
acagttttaa aatccggaaa ggcattgtgt ccaacttccc aaaggccact tatgaagatg 420
tttgacagga tactcggtac acccaatagc gaagcatcac ctgtcaaaaa ggatgcgtct 480
tgaaacacgg acc 493
<210> 2
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
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gcatatcaat aagcggagga aaac 24
<210> 3
<211> 19
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
ggtccgtgtt tcaagacgc 19
Claims (5)
- A 1. Accharomyces cerevisiae, it is characterised in that：Deposit number is CCTCC NO：M2017792.
- 2. application of the saccharomyces cerevisiae described in claim 1 in aquaculture.
- 3. application according to claim 2, it is characterised in that：The saccharomyces cerevisiae is launched into breeding water body to go Except ammonia nitrogen.
- 4. application according to claim 2, it is characterised in that：The additive amount of the saccharomyces cerevisiae is every meter of depth of water per acre Add 200-400g.
- 5. application according to claim 4, it is characterised in that：The living bacteria count of the saccharomyces cerevisiae is 100-150 Hundred million CFU/g.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111377545A (en) * | 2019-07-17 | 2020-07-07 | 环亚(天津)环保科技有限公司 | Water environment body repairing agent for reducing ammonia nitrogen in five types of inferior water |
| CN111676145A (en) * | 2020-06-09 | 2020-09-18 | 青岛玛斯特生物技术有限公司 | Saccharomyces cerevisiae and application thereof in aquaculture |
| CN113881583A (en) * | 2020-07-01 | 2022-01-04 | 安琪酵母股份有限公司 | Strain A-15, breeding method and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111377545A (en) * | 2019-07-17 | 2020-07-07 | 环亚(天津)环保科技有限公司 | Water environment body repairing agent for reducing ammonia nitrogen in five types of inferior water |
| CN111676145A (en) * | 2020-06-09 | 2020-09-18 | 青岛玛斯特生物技术有限公司 | Saccharomyces cerevisiae and application thereof in aquaculture |
| CN111676145B (en) * | 2020-06-09 | 2021-05-28 | 青岛玛斯特生物技术有限公司 | Saccharomyces cerevisiae and application thereof in aquaculture |
| CN113881583A (en) * | 2020-07-01 | 2022-01-04 | 安琪酵母股份有限公司 | Strain A-15, breeding method and application thereof |
| CN113881583B (en) * | 2020-07-01 | 2023-09-12 | 安琪酵母股份有限公司 | Strain An-15, breeding method and application thereof |
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| Publication number | Publication date |
|---|---|
| CN108048338B (en) | 2021-08-13 |
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