CN101864382A - New fermentation method - Google Patents
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- CN101864382A CN101864382A CN201010170295A CN201010170295A CN101864382A CN 101864382 A CN101864382 A CN 101864382A CN 201010170295 A CN201010170295 A CN 201010170295A CN 201010170295 A CN201010170295 A CN 201010170295A CN 101864382 A CN101864382 A CN 101864382A
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Abstract
The invention discloses a new fermentation method, which mainly comprises the following steps of: (1) preparing liquid seeds; (2) preparing a solid plate: adding 18 to 32g of agar into each 1,000 ml of corresponding liquid fermentation culture medium for strict anaerobes, sterilizing, pouring into a plate, cooling, and preparing a solid plate culture medium containing liquid culture medium components by utilizing the solidification capacity of the agar on liquid; and (3) performing amplified fermentation: spraying the corresponding liquid seeds obtained by the step (1) to the surface of the solid plate uniformly, putting the sprayed solid plate in a fermentation chamber at the constant temperature of between 35 and 37 DEG C for closed fermentation culture, culturing for 24 to 72 hours according to different bacterial strains, and collecting bacterial sludge. The method has the advantages of low cost, simple operation, easy subsequent processing and suitability for the production of most of microbial bacterial strains; and through the method, products which break through the upper limit of bacterial concentration in the conventional microbial preparation products greatly can be produced, and about 1,000 billion/g bacterial powder products can be obtained.
Description
Technical field
The invention belongs to the microorganism culturing field, relate to the new fermentation process of a kind of microorganism specifically.
Background technology
At present in the industrial fermentation, the main zymotechnique that adopts completely can classify as two kinds of solid state fermentation and liquid fermentings.
Liquid fermenting be bacterial classification inoculation in fermentor tank, make that somatic cells is free to be suspended in the liquid fermentation medium, and by parameters such as controlled temperature, stirring, air, pressure, nutritive substance concentration, a kind of cultural method that it is grown.The deep-layer liquid cultivation generally needs bubbling air and stirs, and is traditional microbial fermentation cultural method.Mainly there is apparatus expensive in it, and complicated operation, unit substratum produce lower, the more high shortcoming of the isolating cost of bacterium liquid of concentration of thalline quantity.
Solid fermentation comprises two kinds of broad sense and narrow senses, and generalized is: microorganism growth is in water-fast matrix, and contains the free water (free water) of different amounts on the matrix.Narrow sense be: microorganism growth ferments in the water-fast matrix of humidity, in the solid fermentation process, do not contain any free water, along with the increase of the free water of microorganism output, the solid fermentation scope extends to the fermentation that suspends of sticky fermentation (slurry fermentation) and solid particulate.
Solid fermentation method, common process comprises: in the groove jar equipment that is fit to, carry disinfectant, fully soak into the inert support of substratum nutritive ingredient, and microbe inoculation thereon, and, realize a large amount of breedings of target microorganism by controlled temperature and some other correlated condition.Collect thalline or tunning etc. after fermentation is finished, be generally whole collections as product, the step of going forward side by side is carried out post-treatment phase processing such as dry fragmentation.
Solid fermentation mainly has the following advantages:
1. substratum is simple, and such as grain class, Wheat bran, agropyron, large cereal or agricultural-food etc. all can be used fermentation raw material cost less expensive.
2. the matrix pre-treatment is few than liquid fermenting, for example simply adds water and makes the matrix humidity, or simply gall matrix and increase contact area and get final product, does not need specific apparatus, and generally family can carry out step.
3. can produce the special outcome that some liquid fermentings can not produce, the red pigments that produces as red Qu is ten times of liquid fermenting, and Aspergillus has more thermotolerance at the glucosidase that solid fermentation produced than the ferment that liquid fermenting produces again.
4. the recovery purge process in downstream and offal treatment are simplified usually or are simple, often are that whole matrix all is used, and waste is less.
5. but solid fermentation food produces flavour, and improves nutritive value.
At present, though solid fermentation has above advantage, also there is following shortcoming:
1. what the sterilization of material and sterilization were difficult to do is thorough, and cost is also higher, very easily causes living contaminants, influences quality product.
2. be limited to the low wet condition microorganism of growth down,, generally be suitable for fungi so possible flow process and product are more restricted.
3. be difficult for carrying out the uniform distribution of matter and energy, so between yeast phase, the interpolation of material can't reach evenly, and then make ferment effect also inhomogeneous with alr mode.
4. at fine and close environment bottom fermentation, its heat radiation often throws into question, and the often logical hypoxgia in material bottom, and under the uneven situation of stirring, the oxygen consumption bacteria growing is out of condition, especially during mass production, often limits its large-scale production capacity.
5. be not easy to extract pure thalline, usually mix with carrier.
6. subsequent drying technology adopts warm air drying usually, makes number of viable cause greater loss.
7. the bacteria containing amount order of magnitude maximum of finished product generally is no more than about 1,000 hundred million/gram, all is difficult to usually reach about 1,000 hundred million/gram.
Summary of the invention
The technical issues that need to address of the present invention provide a kind of new fermentation process, and this method has raw material and expends minimum, the high advantage of unit substratum product bacterium amount.
The technical scheme that solves the problems of the technologies described above is as follows:
A kind of new fermentation process mainly may further comprise the steps:
(1) seed preparation: carry out conventional bacterial screening and activation earlier, be inoculated in then in the conventional triangular flask liquid nutrient medium, in incubator, cultivate, obtain liquid seeds;
(2) preparation of solid plate: in every 1000ml liquid fermentation medium of the correspondence of non-strictly anaerobic bacterium, add 18-32g agar, pour into after the sterilization in the flat board, utilize the coagulation power of agar, promptly can be made into the solid plate substratum that contains the liquid nutrient medium composition after the cooling liquid;
(3) amplify fermentation: the ferment-seeded of the correspondence that obtains in the step (1) evenly is sprayed on the solid plate surface, be positioned over 35 ℃-37 ℃ indoor sealing fermentation culture of ferment at constant temperature then, difference according to bacterial classification, cultivated 24-72 hour, microorganism will slowly grow into one deck bacterium mud on the plate culture medium surface;
(4) collect bacterium mud: after amplification has been fermented, flat board is taken out proving room, with the hard thin slice (for example ebonite sheet or hard tinsel) of sterilization, the bacterium mud on the plate culture medium is scraped off then, collect, obtain required bacterium mud.
Described fermentation process also comprises: with the bacterium mud that obtains and water by mass ratio 1: 0.8-1.2 is behind stirring and evenly mixing, be bacterium mud by mass ratio again: silicon-dioxide=1: 7.5-8.5 progressively adds dried silica, carry out adsorption dry again, cross 60 mesh sieves, cross 60 mesh sieves after screen overflow pulverized once more, obtain highly enriched pure bacterium powder.
Preferably, described non-strictly anaerobic bacterium is a denitrifying pseudomonas, the consisting of of its solid plate substratum: mainly have among the 1000mL: glucose 4-5g, flour 1.5-2.5g, fish meal 13-17g, saltpetre 4-6g, dipotassium hydrogen phosphate 0.8-1.2g, potassium primary phosphate 0.8-1.2g, sal epsom 0.4-0.6g, yeast extract paste 0.15-0.25g, agar 18-22g.
Preferably, described non-strictly anaerobic bacterium is a subtilis, the consisting of of its solid plate substratum: mainly have among the 1000mL: waste molasses 18-22mL, flour 1.5-2.5g, fish meal 13-17g, liquid protein 9-11mL, manganous sulfate 0.8-1.2g, dipotassium hydrogen phosphate 0.8-1.2g, potassium primary phosphate 0.8-1.2g, sal epsom 0.4-0.6g, yeast extract paste 0.15-0.25g, agar 18-22g.
Preferably, described non-strictly anaerobic bacterium is a Lactobacterium acidophilum, the consisting of of its solid plate substratum: have among the 1000mL: fish meal 18-22g, glucose 18-22g, whey powder 8-12g, agar 28-32g, dipotassium hydrogen phosphate 1.8-2.2g, manganous sulfate: 0.2-0.4g, sodium acetate: 1.5-2.5g, sal epsom: 0.5-0.7g, yeast extract paste: 1.8-2.2g, tween 80 1-2mL.
Fermentation process of the present invention improves by the liquid fermentation medium to routine, makes the solid plate substratum and carries out dull and stereotyped fermentation culture, and it is more suitable in collecting thalline is the fermentative production of main purpose, and has the following advantages:
1, low, simple to operate, the production that can be applicable to most Research for Industrial Microbial Germ (microorganism of non-food product and non-pharmaceutical---non-virulent, non-strictly anaerobic bacterium) of described method cost.
2, can isolate pure bacterium mud easily, it is extremely simple, efficient and cheap that follow-up preservation and complete processing will become.
3, produce the product that the existing microbial preparation product of big quantum jump contains the bacteria concentration upper limit, can arrive 10,000 hundred million/gram left and right sides bacterium powder product.
4, collect the solid medium that is left behind the bacterium mud, not only moisture is lower, and is rich in a large amount of meta-bolitess, can optionally simply be processed into powder, makes an addition in the finished product, or is used for refining extraction meta-bolites.
5, further reduce the discharging of water wasting, refuse, reduce environmental pollution.
Embodiment
Below by specific embodiment, the present invention will be further described, but protection scope of the present invention is not limited by the following examples.
Embodiment 1: the fermentative production of denitrifying pseudomonas
One, the preparation of substratum
1, seed culture medium (by 1000mL)
Glucose 10g, dipotassium hydrogen phosphate: 1g, potassium primary phosphate: 1g, Trisodium Citrate: 10g, saltpetre: 5g, sal epsom: 0.2g, yeast extract paste: 1g, adding water to cumulative volume is 1000mL, pH transfers to 8.
With the packing of 500mL triangular flask, each triangular flask packing 400mL liquid; Sterilization 40min.
2, the solid plate of fermention medium (by 1000mL)
Glucose 5g, flour 2g, fish meal 15g, saltpetre 5g, dipotassium hydrogen phosphate 1g, potassium primary phosphate 1g, sal epsom 0.5g, yeast extract paste 0.2g, agar 20g, adding water to cumulative volume is 1000mL, pH transfers to 8.0.
Fish meal is handled: fish meal is with quantitative water boil and continue 20～30min, and the amount of water need calculate the substratum the inside; Treated fish meal is with 40 purpose screen filtrations, and filtrate directly is added to substratum.All the other indissoluble composition additional heat may dissolving backs add in the substratum, and fermention medium is promptly joined i.e. sterilization and promptly used; 121 ℃ of sterilization 20min.
Preparation solid plate substratum:
Stainless steel plate is cleaned, dried,, cool off stand-by 110 ℃ of dry sterilizations of baking oven 1 hour.This step was finished in inoculation in preceding 2 hours.
Fermention medium is promptly joined the i.e. usefulness of promptly sterilizing, and after having sterilized, the rear plate of will sterilizing lies against on aseptic processing room's desktop, toward the dull and stereotyped substratum that quantitatively injects about 250mL, treats promptly can inoculate after the agar cooled and solidified then.Every liter of fermention medium can prepare 4 flat boards approximately.
Two, fermentative production:
1. seed preparation
Aseptic technique after bacterial classification activated, is got 2～3 ring bacterium mud and is inserted in the seed culture medium triangular flasks, and 37 ℃ of constant temperature shaking table middling speeds are cultivated 24～30h, seed liquor.
The feature of certified seed: bacterium liquid turbidity is higher, and the thalline branch is evenly distributed, and obvious aerogenesis is arranged; During microscopy (carbolfuchsin dyeing), thalline is in vigorous division stage, and the form unanimity shows as tiny bacillus, and thalline is long slightly, and the division node is many, does not have obviously assorted bacterium.
2. solid plate fermentation
Set up one or more ferment at constant temperature chamber, each about 4-8 square metre big, and about 2.5 meters high, stopping property and thermal insulation are good, and interior dress UV-lamp and fumigator are convenient to sterilization.Suitably big or small multilayer stainless steel weldering frame of indoor placement is used to stack the stainless steel pallet.To set up an aseptic processing room in addition, the operation of operation before being used to ferment.
Seed liquor evenly is sprayed on above-mentioned each piece solid plate substratum, shakes gently to seed liquor and be paved with planar surface, (100 Stainless Steel Discs need about the 400ml seed liquor altogether).Be inverted laminated flat then in 37 ℃ of cultivations of culturing room's constant temperature.Beginning is observed flat board every 12h, and the thalli growth situation is carried out record.Every 3h microscopy once after the 12h.About the 24h of inoculation back, denitrifying bacteria generates red bacterium mud at planar surface, and after microscopy was qualified, can collect this moment.
3, microorganism collection
Take out dull and stereotypedly, the bacterium mud on the flat board is scraped off, collect in the vessel such as beaker with the hard sheet or the tinsel that disinfect.1: 1 in proportion (mass ratio) abundant stirring and evenly mixing in the miniature liquid stirrer of good bacterium mud and water will be collected, stir behind the 20min bacterium mud in proportion: water: silicon-dioxide=1: 1: 8 (mass ratio) progressively adds dried silica, in having the stirrer of vibratory screening apparatus, carry out adsorption dry, simultaneously continuous stirring and evenly mixing and sieve (60 order), sieve once more after screen overflow pulverized, can form highly enriched pure bacterium powder, this moment, moisture content was about 10%～15%, bacterial content can reach 10,000,000,000,000/gram, add 0.5% calcium propionate as mould inhibitor after, salable normal temperature is placed for a long time.This bacterium powder can further dilute makes the finished product.
Embodiment 2: the dull and stereotyped fermentation manufacturing technique of subtilis
One, the preparation of substratum
1, seed culture medium (by 1000mL)
Extractum carnis 3g, peptone 10g, sodium-chlor 5g, glucose 10g, add water to 1000mL, regulate pH8.
With the packing of 500mL triangular flask, each triangular flask packing 250mL liquid; Sterilization 30min.
2, fermention medium (by 1000mL)
Waste molasses 20mL, flour 2g, fish meal 15g, liquid protein 10mL, manganous sulfate 1g, dipotassium hydrogen phosphate 1g potassium primary phosphate 1g, sal epsom 0.5g, yeast extract paste 0.2g, agar 20g add water to 1000ml, and pH transfers to 8.The processing of fish meal and the preparation of plate culture medium such as embodiment 1.
Two, fermentative production:
1. seed preparation
Aseptic technique after bacterial classification activated, is got 2～3 ring bacterium mud and is inserted in the seed culture medium triangular flasks, and 37 ℃ of constant temperature shaking table middling speeds are cultivated 24～30h.
The feature of certified seed: bacterium liquid turbidity is higher, and mycoderm is less, and the thalline branch is evenly distributed; During microscopy (carbolfuchsin dyeing), thalline is in vigorous division stage, and the form unanimity can be seen based on cenobium, shows as thick stock bacterium, and thalline is very long, and the division node is many, does not have obviously assorted bacterium.Be the best inoculation fermentation time this moment.
2. solid plate fermentation
Seed liquor evenly is sprayed on above-mentioned each piece solid plate substratum, shakes gently to seed liquor and be paved with planar surface, (100 Stainless Steel Discs need about the 400ml seed liquor altogether).Be inverted laminated flat then in 37 ℃ of cultivations of culturing room's constant temperature.
Beginning is observed flat board every 12h, and the thalli growth situation is carried out record.Every 3h microscopy once after the 12h.
Behind the inoculation 6h, planar surface begins to see a small amount of bacterium colony, is evenly distributed, and slight genus bacillus smell is arranged; Can see during microscopy thalline shorten chap (with the length of liquid seeds relatively), divide quite vigorously, cenobium is more, incubator is opened in the pollution of the bacterium of should noting this moment mixing as far as possible less.
Behind the inoculation 12h, this moment, the genus bacillus cell concentration almost reached equilibrium state, and also near low-level, thalline begins to produce gemma in moisture content and nutritive substance consumption simultaneously; Planar surface can white mycoderm (bacterium mud) obviously occur, and the mycoderm surface ratio is more coarse, is evenly distributed, and does not have obviously outstanding single bacterium colony, and wetting ability is relatively poor, and the globule can not permeate on the mycoderm surface, and incubator has the smell of stronger genus bacillus; During the microscopy sampling, the mycoderm ratio is easier to break away from substratum, and viscosity is undesired, and assorted bacterium is more; During 100 * oily sem observation, thalline begins to stop division, and the thalline degree of scatter is higher.Produced gemma in the thalline possibility this moment, if there is independent gemma to distribute, then needs to calculate the gemma rate; More than the gemma rate reached 7 one-tenth, perhaps thalline (it is opaque fully to dye) can be collected bacterium mud more at least, need not wait until next step.
15h is to 18h in inoculation, and this moment, the gemma parent separated with gemma basically, and gemma independently exists, and decides to collect immediately bacterium mud on practical situation.
Attention: the major reason that makes genus bacillus produce gemma is: the minimizing of moisture.Therefore should collect bacterium mud when producing than plurispore, therefore overlong time can grow the bacterium of mixing, and the powerful interior moisture of solid medium that absorbs of part genus bacillus growth simultaneously causes substratum over-drying, influences the collection of bacterium mud.
3. microorganism collection
Step can obtain highly enriched pure bacterium powder with embodiment 1, and this moment, moisture content was about 10%～15%, and bacterial content can reach 10,000,000,000,000/gram, add 0.5% calcium propionate as mould inhibitor after, salable normal temperature is placed for a long time.This bacterium powder can further dilute makes the finished product.
Embodiment 3: the dull and stereotyped fermentation manufacturing technique of Lactobacterium acidophilum
One, the preparation of substratum
1, seed culture medium (by 1000mL)
Extractum carnis 10g, peptone 10g, whey powder 10g, glucose 20g, dipotassium hydrogen phosphate: 2g, manganous sulfate: 0.3g, dibasic ammonium citrate: 10g, sodium acetate: 2g, sal epsom: 0.6g, yeast extract paste: 5g, tween 80 1mL, add water to 1000mL, pH transfers to 6.2～6.4.
With the packing of 500mL triangular flask, each triangular flask packing 300mL liquid; Sterilization 20min.
2, fermention medium (by 1000mL)
Fish meal 20g, glucose 20g, whey powder 10g, agar 30g, dipotassium hydrogen phosphate: 2g, manganous sulfate: 0.3g, sodium acetate: 2g, sal epsom: 0.6g, yeast extract paste: 2g, tween 80 1-2mL add water to 1000mL pH and transfer to 6.2～6.4.
The processing of fish meal and the preparation of plate culture medium such as embodiment 1.
Two, fermentative production
1. seed preparation
Aseptic technique after bacterial classification activated, is got 2～3 ring bacterium mud and is inserted in the seed culture medium triangular flasks, and 37 ℃ of constant temperature shaking table middling speeds are cultivated 24～30h, seed liquor.
The feature of certified seed: bacterium liquid turbidity is higher, and the thalline branch is evenly distributed, and pH obviously descends, and produces tangible sour fragrance flavor; During microscopy (carbolfuchsin dyeing), thalline is in vigorous division stage, and the form unanimity shows as medium sized bacillus, two circle section, and thalline is thicker, does not have obviously assorted bacterium.
2. solid plate fermentation
Seed liquor evenly is sprayed on above-mentioned each piece solid plate substratum, shakes gently to seed liquor and be paved with planar surface, (100 Stainless Steel Discs need about the 400ml seed liquor altogether).Be inverted laminated flat then in 37 ℃ of cultivations of culturing room's constant temperature.
Beginning is observed flat board every 12h, and the thalli growth situation is carried out record.Every 3h microscopy once after the 12h.
About the 48h of inoculation back, Lactobacterium acidophilum has generated the bacterium mud of white at planar surface, and after microscopy was qualified, can collect this moment.
3. microorganism collection
Step obtains highly enriched pure bacterium powder with embodiment 1, and this moment, moisture content was about 10%～15%, and bacterial content can reach 10,000,000,000,000/gram, add 0.5% calcium propionate as mould inhibitor after, salable normal temperature is placed for a long time.This bacterium powder can further dilute makes the finished product.
More than be at the specifying of possible embodiments of the present invention, but this embodiment is not in order to limiting claim of the present invention, does not allly break away from equivalence of the present invention and implement or change, all should be contained in the claim of the present invention.
Claims (5)
1. a new fermentation process is characterized in that, mainly may further comprise the steps:
(1) seed preparation: carry out conventional bacterial screening and activation earlier, be inoculated in then in the conventional triangular flask liquid nutrient medium, in incubator, cultivate, obtain liquid seeds;
(2) preparation of solid plate: in every 1000ml liquid fermentation medium of the correspondence of non-strictly anaerobic bacterium, add 18-32g agar, pour into after the sterilization in the flat board, promptly can be made into the solid plate substratum that contains the liquid nutrient medium composition after the cooling;
(3) amplify fermentation: the liquid seeds of the correspondence that obtains in the step (1) evenly is sprayed on the solid plate media surface, is positioned over 35 ℃-37 ℃ indoor sealing fermentation culture of ferment at constant temperature then,, cultivated 24-72 hour according to the difference of bacterial classification;
(4) collect bacterium mud: after amplification has been fermented, flat board is taken out proving room, with the hard thin slice of sterilization, the bacterium mud on the plate culture medium is scraped off then, collect, obtain required bacterium mud.
2. fermentation process according to claim 1, it is characterized in that, described fermentation process also comprises: resulting bacterium mud of step (4) and water are pressed mass ratio 1: 0.8-1.2 is behind stirring and evenly mixing, be bacterium mud by mass ratio again: after silicon-dioxide=1: 7.5-8.5 progressively adds dried silica, carry out adsorption dry, cross 60 mesh sieves, cross 60 mesh sieves after screen overflow is pulverized once more, obtain highly enriched pure bacterium powder.
3. fermentation process according to claim 1 and 2 is characterized in that, described non-strictly anaerobic bacterium is a denitrifying pseudomonas, the consisting of of its solid plate substratum:
Mainly have among the 1000mL: glucose 4-5g, flour 1.5-2.5g, fish meal 13-17g, saltpetre 4-6g, dipotassium hydrogen phosphate 0.8-1.2g, potassium primary phosphate 0.8-1.2g, sal epsom 0.4-0.6g, yeast extract paste 0.15-0.25g, agar 18-22g.
4. fermentation process according to claim 1 and 2 is characterized in that, described non-strictly anaerobic bacterium is a subtilis, the consisting of of its solid plate substratum:
Mainly have among the 1000mL: waste molasses 18-22mL, flour 1.5-2.5g, fish meal 13-17g, liquid protein 9-11mL, manganous sulfate 0.8-1.2g, dipotassium hydrogen phosphate 0.8-1.2g, potassium primary phosphate 0.8-1.2g, sal epsom 0.4-0.6g, yeast extract paste 0.15-0.25g, agar 18-22g.
5. fermentation process according to claim 1 and 2 is characterized in that, described non-strictly anaerobic bacterium is a Lactobacterium acidophilum, the consisting of of its solid plate substratum:
Have among the 1000mL: fish meal 18-22g, glucose 18-22g, whey powder 8-12g, agar 28-32g, dipotassium hydrogen phosphate 1.8-2.2g, manganous sulfate: 0.2-0.4g, sodium acetate: 1.5-2.5g, sal epsom: 0.5-0.7g, yeast extract paste: 1.8-2.2g, tween 80 1-2mL.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105441360A (en) * | 2015-12-27 | 2016-03-30 | 陈景河 | Organic fertilizer fermentation inoculant |
| CN105695373A (en) * | 2016-04-18 | 2016-06-22 | 广州市佰沃生物科技有限公司 | Bacillus subtilis(natto) fermentation method |
| CN106007865A (en) * | 2016-06-27 | 2016-10-12 | 安徽省蚌埠市中远肥业有限公司 | Organic fertilizer special for rice in tillering stage and preparation method of organic fertilizer |
| CN106116764A (en) * | 2016-06-27 | 2016-11-16 | 安徽省蚌埠市中远肥业有限公司 | A kind of single cropping Caulis Zizaniae caduciflorae fertilizer special for organic and preparation method thereof |
| CN106116959A (en) * | 2016-06-27 | 2016-11-16 | 安徽省蚌埠市中远肥业有限公司 | A kind of rice seedling phase fertilizer special for organic and preparation method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105441360A (en) * | 2015-12-27 | 2016-03-30 | 陈景河 | Organic fertilizer fermentation inoculant |
| CN105695373A (en) * | 2016-04-18 | 2016-06-22 | 广州市佰沃生物科技有限公司 | Bacillus subtilis(natto) fermentation method |
| CN105695373B (en) * | 2016-04-18 | 2019-05-24 | 广州市佰沃生物科技有限公司 | The fermentation process of bafillus natto |
| CN106007865A (en) * | 2016-06-27 | 2016-10-12 | 安徽省蚌埠市中远肥业有限公司 | Organic fertilizer special for rice in tillering stage and preparation method of organic fertilizer |
| CN106116764A (en) * | 2016-06-27 | 2016-11-16 | 安徽省蚌埠市中远肥业有限公司 | A kind of single cropping Caulis Zizaniae caduciflorae fertilizer special for organic and preparation method thereof |
| CN106116959A (en) * | 2016-06-27 | 2016-11-16 | 安徽省蚌埠市中远肥业有限公司 | A kind of rice seedling phase fertilizer special for organic and preparation method thereof |
| CN106116868A (en) * | 2016-06-27 | 2016-11-16 | 安徽省蚌埠市中远肥业有限公司 | A kind of Rice during Grain Filling Stage fertilizer special for organic and preparation method thereof |
| CN106187571A (en) * | 2016-06-27 | 2016-12-07 | 安徽省蚌埠市中远肥业有限公司 | A kind of Oryza sativa L. long fringe phase fertilizer special for organic and preparation method thereof |
| CN112179091A (en) * | 2020-10-16 | 2021-01-05 | 安徽沃园生物科技有限公司 | A even drying device for fertilizer |
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