CN105613707A - Bacillus subtilis biological preservative and application thereof to preservation of large yellow croakers - Google Patents
Bacillus subtilis biological preservative and application thereof to preservation of large yellow croakers Download PDFInfo
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- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 46
- 235000014469 Bacillus subtilis Nutrition 0.000 title claims abstract description 43
- 238000004321 preservation Methods 0.000 title claims abstract description 24
- 239000003755 preservative agent Substances 0.000 title claims abstract description 17
- 230000002335 preservative effect Effects 0.000 title abstract description 9
- 241001596950 Larimichthys crocea Species 0.000 title abstract description 6
- 229920001661 Chitosan Polymers 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 11
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- 238000000855 fermentation Methods 0.000 claims description 17
- 230000004151 fermentation Effects 0.000 claims description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 239000000287 crude extract Substances 0.000 claims description 12
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- 239000001963 growth medium Substances 0.000 claims description 7
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- 241000589516 Pseudomonas Species 0.000 description 2
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- 239000013543 active substance Substances 0.000 description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 2
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- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
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- 238000005265 energy consumption Methods 0.000 description 1
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- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
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- 239000004467 fishmeal Substances 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
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- 239000008273 gelatin Substances 0.000 description 1
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- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
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- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
- A23B4/08—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block
- A23B4/09—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block with direct contact between the food and the chemical, e.g. liquid N2, at cryogenic temperature
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides application of bacillus subtilis to preservation of large yellow croakers as a biological preservative. The bacillus subtilis is bacillus subtilis BSh0851. The invention further provides a preservation method of the large yellow croakers; the fresh large yellow croakers are inactivated by crushed ice, and bacillus subtilis rough extract, nisaplin and a chitosan solution are mixed and then are sprayed on fish bodies; and then the large yellow croakers are stored on ice in a refrigerator at 4 DEG C. The method has a simple and safe process; the preservation and fresh-keeping effects are remarkable; and the shelf life of aquatic products is effectively prolonged, and original freshness, flavor and texture of the aquatic products can be kept very well.
Description
Technical field
The invention belongs to aquatic products antiseptic preservation technology, be specifically related to the bacillus subtilis method as bio-preservative application in Carnis Pseudosciaenae anti-corrosive fresh-keeping and preservation and antisepsis.
Background technology
Aquatic products are the important component parts of people's food, and chilled preservation is Aquatic product at one of major way of market circulation. Contain the nutrient substance such as rich in protein due to aquatic products, it is easy to causing Putrefying bacteria fast-growth to breed, corrupt substance accumulates in a large number, thus very easily rotten, has influence on the quality of aquatic products, even threaten that people's is healthy. Therefore, the preservation method of aquatic products becomes current study hotspot. At present, the preservation technique being applied to aquatic products mainly has preservation by low temperature, fresh chemically, gas preservation, radiation preservation and biological preservation etc. The advantage that biological preservation is high with its safety, energy consumption is low becomes the new focus of exploitation. Plant source preservative acts primarily as antioxidation; Animal sources antibacterial such as protamine; Microbial source antibacterial conventional in aquatic products mainly has the lactein that lactic acid bacteria produces, its mycin etc. of receiving. The antistaling agent of dynamic/plant origin is limited by raw material, and price is higher; And the microbial source antistaling agent of industrialization at present is microbe metabolite, it is increasingly subject to pay attention to.
Bacillus subtilis strain has the activity suppressing pathogen and microalgae, is allowed to use in fruit and vegerable, fish meal.
Summary of the invention
It is an object of the invention to provide a kind of bacillus subtilis as bio-preservative application in preservation of fishery anticorrosion, for the approach that the fresh-keeping offer of aquatic products particularly chilled Carnis Pseudosciaenae is new.
For achieving the above object, the present invention adopts the following technical scheme that
Bacillus subtilis described in this application is bacillus subtilis (Bacillussubtulis) BSh0851, preservation date is December in 2009 13, depositary institution is China Microbiological bacterial strain preservation administration committee's common micro-organisms center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, preserving number is CGMCCNo.3502.
Described bacillus subtilis (Bacillussubtilis) is gram positive bacteria, and spore ellipse, to column, is positioned at thalline central or slightly inclined, and after sporulation, thalline does not expand, and bacterium colony rough surface is opaque, dirty white or slightly yellow, is commonly formed wrinkle mould; Aerobe, available protein, multiple sugar and starch, V-P is positive.
Adopt the condition of culture after optimization culture, prepare the fermentation liquid containing high active substance by liquid fermentation, and adopt the method for Acid precipitation to obtain lipopeptid class active substance crude extract. Crude extract yield is 0.45g/L, and the bacteriostasis rate to 105cfu/mL Psychrobacter, 105cfu/mL pseudomonas and 105cfu/mL Shewanella 3 bacterium mixed liquor is 33%.
By 0.3mg/mL crude extract and the general woods of 0.3mg/mL Nysa and 20mg/mL chitosan hydrochlorate compatibility, make mixing antistaling agent. Carnis Pseudosciaenae is carried out fresh-keeping by the antistaling agent using the present invention, after storing 5 days, total volatile basic nitrogen (TVB-N) and microorganism and organoleptic indicator all meet one-level freshness in the related sanitary standards (SC/T3101-2010) such as China fresh Radix et Rhizoma Rhei fish, corrosion-resistanting fresh-keeping effect is notable, extends the shelf life of Carnis Pseudosciaenae. Result shows that fermentation of bacillus subtilis liquid crude extract can suppress the growth of Carnis Pseudosciaenae surface bacteria effectively, and also have some improvement to the fatty acid oxidation of Pseudosciaena crocea meat and freshness effect simultaneously.
Carnis Pseudosciaenae preservation method is to be inactivated by fresh and alive Carnis Pseudosciaenae with trash ice, bacillus subtilis crude extract, the general woods of Nysa, the mixing of chitosan hydrochlorate solution is sprayed at fish body, preserves on ice in 4 DEG C of refrigerators.
Specifically include following steps: prepared by (1) seed liquor: the bacillus subtilis that picking one inoculating loop has activated from nutrient agar slant medium is inoculated in seed culture medium, 37 DEG C, 200r/min, cultivate 24h; Seed culture medium is Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, peptone 10g, distilled water 1000mL, pH7.0-7.2;
(2) fermentation liquid medium preparing: yeast extract 14g, anhydrous glucose 14g, MgSO41g, distilled water 1000mL, pH7.0;
(3) bacillus subtilis is without the preparation of thalline fermentation liquid: accessing the seed liquor that step (1) prepares in the fermentation liquid that step (2) prepares, inoculum concentration is 7%; Regulate original ph 8; Cultivation temperature 38 DEG C, rotating speed 100r min-1, liquid amount 150mL/250mL shaking flask; Cultivate 24h with this understanding;
(4) without the preparation of fermented liquid: by described for step (3) bacillus subtilis bacteria culture fluid after the centrifugal 15min of 10000r/min, with 0.22 ��m of membrane filtration, be the ferment filtrate without thalline;
(5) prepared by fermentation broth coarse extract: described for step (4) HCl without fermented liquid 2mol/L is adjusted to pH2.0, places 3d in 4 DEG C, and the centrifugal 10min of 10000r/min, collects precipitate, be dissolved in sterilized water again, and regulates pH to 7 with NaCl;
(6) yield obtaining Substance and antibacterial fat peptide matters is sunk in the fermentation liquor acid of step (5) gained is 0.45g/L, to 105Cfu/mL Psychrobacter, 105Cfu/mL pseudomonas and 105The bacteriostasis rate of cfu/mL Shewanella 3 bacterium mixed liquor is 33%;
(7) diluting the crude extract of step (5) for active matter quality is 0.3mg/mL, makes mix antistaling agent with the general woods of 0.3mg/mL Nysa and 20mg/mL chitosan hydrochlorate;
(8) on the full surface of chilled Carnis Pseudosciaenae, spraying, every 12h, the mixing antistaling agent that step (7) prepares, the consumption that every tail Carnis Pseudosciaenae is sprayed every time is about 5mL.
Beneficial effects of the present invention is as follows: 1, bacillus subtilis is as a kind of feedstuff probiotic bacteria, and its metabolite adds safe to the human body harmless. 2, the antimicrobial spectrum of this bacillus subtilis crude extract is wider, it is possible to the effective growth suppressing Gram-positive and gram negative bacteria, and preservation has obvious action under 4 DEG C of conditions, thus the anti-corrosive fresh-keeping of Carnis Pseudosciaenae is played good effect. 3, this bacillus subtilis source composite preservative can effectively keep the freshness of aquatic products, sensory properties etc. in the process to preservation of fishery such as Carnis Pseudosciaenae.
Detailed description of the invention
The present invention is with total volatile basic nitrogen (TVB-N), and thiobarbituricacid��-(TBA), the metrics evaluation bacillus subtilis source composite preservative such as total plate count is to cultivating chilled Carnis Pseudosciaenae fresh-keeping effect.
Method with reference to the mensuration total volatile basic nitrogen that the bioassay standard (SC/T3032-2007) of total volatile basic nitrogen in fishery products specifies. 10g(is claimed to be accurate to 0.01g) in homogenizing cup, add 90mL perchloric acid solution, homogenizing 2min, with filter paper filtering or centrifugation, filtrate, in the storage under ambient conditions of 2 DEG C-6 DEG C, can preserve 2d. Pour the boric acid of 10mL2% into 50mL conical flask, add 1-2 and drip mixed indicator, boric acid is placed in condensing tube lower end, and make condensing tube lower end immersed in liquid level, accurately draw above-mentioned sample filtrate 2mL in distillator reacts, add 1% magnesium oxide suspension 5mL, rapid capping plug, add water, pass into steam. Treating that steam is full of inside distillator, condensing tube calculate there is first condensed water, namely distillation 5min stops, and absorbs liquid about 0.01mol/L hydrochloric acid standard solution titration. Terminal takes on a red color, and does reagent blank test simultaneously. TVB-N value unit is mgN/100g. Total volatile basic nitrogen (TVBN) is one of primary chemical index judging Fish freshness, and the TVBN value of fish one-level freshness is at 5-10mg/100g, and the fish TVBN value of general freshness is at 15-25mg/100g, and when TVBN value, to reach 30-40mg/100g then fish corrupt.
Total plate count assay method: the total plate count in the safe national standard-food microbiological examination of reference food measures operation (GB4789.2-2010). Fresh marine fish bacteria colony count is made regulation by national standard GB/T 18108-2008: total number of bacteria��10 in the flesh of fish4Cfu/g is one-level freshness (A level) standard ,��105Cfu/g standard is corrupt terminal (C level) standard.
Divide 4 groups by fresh Carnis Pseudosciaenae, be respectively adopted chitosan hydrochlorate, bacillus subtilis crude extract-chitosan hydrochlorate and bacillus subtilis crude extract-Nysa Pu Lin-chitosan hydrochlorate composite preservative and carry out fresh-keeping; And do 1 group of comparison, in order to ensure the concordance of experimental factors, matched group sprays sterilized water.
(1) prepared by seed liquor: the bacillus subtilis that picking one inoculating loop has activated from nutrient agar slant medium is inoculated in seed culture medium, 37 DEG C, 200r/min, cultivates 24h; Seed culture medium is Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, peptone 10g, distilled water 1000mL, pH7.0-7.2;
(2) fermentation liquid medium preparing: yeast extract 14g, anhydrous glucose 14g, MgSO41g, distilled water 1000mL, pH7.0;
(3) bacillus subtilis is without the preparation of thalline fermentation liquid: accessing the seed liquor that step (1) prepares in the fermentation liquid that step (2) prepares, inoculum concentration is 7%; Regulate original ph 8; Cultivation temperature 38 DEG C, rotating speed 100r min-1, liquid amount 150mL/250mL shaking flask; Cultivate 24h with this understanding;
(4) without the preparation of fermented liquid: by described for step (3) bacillus subtilis bacteria culture fluid after the centrifugal 15min of 10000r/min, with 0.22 ��m of membrane filtration, be the ferment filtrate without thalline;
(5) prepared by fermentation broth coarse extract: described for step (4) HCl without fermented liquid 2mol/L is adjusted to pH2.0, places 3d in 4 DEG C, again the centrifugal 10min of 10000r/min, collect precipitate, it is dissolved in sterilized water, and regulates pH to 7 with NaCl, it is thus achieved that bacillus subtilis crude extract;
Diluting bacillus subtilis crude extract for active matter quality is 0.3mg/mL, makes mix antistaling agent with the general woods of 0.3mg/mL Nysa and 20mg/mL chitosan hydrochlorate.
Embodiment 1
Bacillus subtilis crude extract-Nysa Pu Lin-chitosan hydrochlorate composite preservative is adopted to carry out fresh-keeping. Chilled Carnis Pseudosciaenae positive and negative is uniformly sprayed the antistaling agent of about 10mL.
Storage: Carnis Pseudosciaenae is preserved on ice at 4 DEG C. After preserving 5 days (d), total plate count is more than 105Cfu/g, 6 days (d) total volatile basic nitrogen afterwards (TVB-N) content is more than 25mg/100g, and shelf life terminates.
Embodiment 2
Bacillus subtilis crude extract-chitosan hydrochlorate composite preservative is adopted to carry out fresh-keeping. Chilled Carnis Pseudosciaenae positive and negative is uniformly sprayed the antistaling agent of about 10mL.
Storage: Carnis Pseudosciaenae is preserved on ice at 4 DEG C. After preserving 3.5 days (d), total plate count is more than 105Cfu/g, 4 days (d) total volatile basic nitrogen afterwards (TVB-N) content is more than 25mg/100g, and shelf life terminates.
Embodiment 3
Chitosan hydrochlorate composite preservative is adopted to carry out fresh-keeping. Chilled Carnis Pseudosciaenae positive and negative is uniformly sprayed the antistaling agent of about 10mL.
Storage: Carnis Pseudosciaenae is preserved on ice at 4 DEG C. After preserving 2.5 days (d), total plate count is more than 105Cfu/g, 3 days (d) total volatile basic nitrogen afterwards (TVB-N) content is more than 25mg/100g, and shelf life terminates.
Embodiment 4(is than embodiment) chilled Carnis Pseudosciaenae positive and negative is uniformly sprayed the sterilized water of about 10mL. Storage: Carnis Pseudosciaenae is preserved on ice at 4 DEG C. After preserving 2 days (d), total plate count is more than 105Cfu/g, and total volatile basic nitrogen (TVB-N) content is more than 25mg/100g, shelf life terminates.
Embodiment 5
The separation of bacillus subtilis and qualification
(1) this strains separation is in Apis body. From beehive, take apis mellifera worker bee adult 30 at random, be placed in aseptic centrifuge tube. Under sterile working, add the alcohol-pickled 5s of 70%, take out polypide and be placed in 0.1% mercuric chloride liquid sterilization 2min, move to and aquesterilisa is changed clothes 3 times, then polypide is placed in aseptic centrifuge tube, adds the sterilized water of 5mL, concussion, take 50 �� L and coat in culture medium, check that whether body surface sterilizing is complete. Honeybee body after sterilization is placed in the cake wax of sterilization on superclean bench and dissects. Take out its intestinal in aseptic mortar, add the grinding of 1mL sterilized water, lapping liquid sterilized water is diluted to 10-1-10-6Each dilution factor takes 50 �� L spread plates and is easily separated, and each dilution factor repeats 3 times, after cultivating 72h in the constant incubator of 30 DEG C, labelling characterizes different bacterium colonies so that purification is cultivated, and by countability principle, bacterium colonies different on each flat board is counted. Picking characterizes different single bacterium colony antibacterial line purification, obtains pure culture, inclined-plane conservation. Choose colony growth dilution factor sparse, suitable and calculate clump count, obtain 3 meansigma methodss repeated. Calculate the bacterial population in every gram of (milliliter) intestinal. Bacterial population (colony-forming units)=flat board in every gram of (mL) intestinal adds on clump count �� extension rate/flat board the amount (mL) of bacterium solution.
(2) qualification of Apis intestinal bacteria
With reference to " common bacteria system identification handbook " (the elegant pearl in east; Cai Miaoying, 2001) and " uncle's Jie Shi Bacteria Identification handbook " (1984) the 8th editions experimental techniques, carry out Gram��s staining, flagella staining, spore staining, glucose oxidative fermentation, catalase, indole, V-P, M.R, gelatin liquefaction, produce H2S, phenylalanine, deaminase, the experiments such as citrate, malonate, sucrose, lactose and mannose utilize, nitrate reduction carry out adhering to separately of each section, and each genus identification mark is described.
Identify that the purpose bacterial strain obtaining separating purification is bacillus subtilis (Bacillussubtilis). Gram positive bacteria, spore ellipse, to column, is positioned at thalline central or slightly inclined, and after sporulation, thalline does not expand, and bacterium colony rough surface is opaque, dirty white or slightly yellow, is commonly formed wrinkle mould; Aerobe, available protein, multiple sugar and starch, V-P is positive.
The foregoing is only presently preferred embodiments of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to the covering scope of the present invention.
Claims (6)
1. a bacillus subtilis, it is characterized in that: described bacterial strain is bacillus subtilis BSh0851, preservation date is December in 2009 13, and depositary institution is China Microbiological bacterial strain preservation administration committee's common micro-organisms center, and preserving number is CGMCCNo.3502.
2. a bacillus subtilis as claimed in claim 1 application in preparing bio-preservative.
3. bacillus subtilis bio-preservative according to claim 2, it is characterized in that: described antistaling agent is the dilution of bacillus subtilis crude extract is 0.3mg/mL for active matter quality, makes mix antistaling agent with the general woods of 0.3mg/mL Nysa and 20mg/mL chitosan hydrochlorate.
4. bacillus subtilis bio-preservative according to claim 2, it is characterised in that: the preparation method of bacillus subtilis crude extract is:
(1) prepared by seed liquor: the bacillus subtilis that picking one inoculating loop has activated from nutrient agar slant medium is inoculated in seed culture medium, 37 DEG C, 200r/min, cultivates 24h; Seed culture medium is Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, peptone 10g, distilled water 1000mL, pH7.0-7.2;
(2) fermentation liquid medium preparing: yeast extract 14g, anhydrous glucose 14g, MgSO41g, distilled water 1000mL, pH7.0;
(3) bacillus subtilis is without the preparation of thalline fermentation liquid: accessing the seed liquor that step (1) prepares in the fermentation liquid that step (2) prepares, inoculum concentration is 7%; Regulate original ph 8; Cultivation temperature 38 DEG C, rotating speed 100r min-1, liquid amount 150mL/250mL shaking flask; Cultivate 24h with this understanding;
(4) without the preparation of fermented liquid: by described for step (3) bacillus subtilis bacteria culture fluid after the centrifugal 15min of 10000r/min, with 0.22 ��m of membrane filtration, be the ferment filtrate without thalline;
(5) prepared by fermentation broth coarse extract: described for step (4) HCl without fermented liquid 2mol/L is adjusted to pH2.0, places 3d in 4 DEG C, and the centrifugal 10min of 10000r/min, collects precipitate, be dissolved in sterilized water again, and regulates pH to 7 with NaCl.
5. bacillus subtilis bio-preservative application in Carnis Pseudosciaenae anti-corrosive fresh-keeping as claimed in claim 2.
6. application according to claim 2, it is characterized in that: Carnis Pseudosciaenae preservation method is to be inactivated by fresh and alive Carnis Pseudosciaenae with trash ice, bacillus subtilis crude extract, the general woods of Nysa, the mixing of chitosan hydrochlorate solution are sprayed at fish body, spraying described bio-preservative every 12h, the consumption that every tail Carnis Pseudosciaenae is sprayed every time is 5-10mL; Preserve on ice in 4 DEG C of refrigerators.
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| CN107760308A (en) * | 2017-10-18 | 2018-03-06 | 福州大学 | A kind of soil conditioner containing bacillus subtilis and chitosan |
| CN107760308B (en) * | 2017-10-18 | 2019-06-07 | 福州大学 | A kind of soil conditioner containing bacillus subtilis and chitosan |
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| CN109456922A (en) * | 2018-11-30 | 2019-03-12 | 江苏大学 | The beach the Halophilic Bacterium bacterial strain Hua Jin bacillus of one plant of raising alec fermentation quality |
| CN109456922B (en) * | 2018-11-30 | 2021-09-10 | 江苏大学 | Moderately halophilic bacteria strain Zhan beach bacillus for improving fermentation quality of fish paste |
| CN114128754A (en) * | 2021-05-18 | 2022-03-04 | 郭云 | Bacillus subtilis antistaling agent and preparation method thereof |
| RU2797684C1 (en) * | 2022-08-23 | 2023-06-07 | Ирина Васильевна Ткачева | Method for keeping live fish during transportation and storage |
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