CN102515952A - Chitosan coated alga bioorganic fertilizer and preparation method thereof - Google Patents
Chitosan coated alga bioorganic fertilizer and preparation method thereof Download PDFInfo
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Abstract
The invention provides a chitosan coated alga bioorganic fertilizer and a preparation method thereof. The chitosan coated alga bioorganic fertilizer is obtained by adopting a process combining high efficiency physical fragmentation with a chemical processing technology to extract physiological active substances and nutrients of algae, adding zymocytes with the synergism to alga residues, preparing an alga organic fertilizer in an aerobic fermentation mode, adding proper amounts of nitrogen phosphorous potassium, trance elements and functional bacteria, mixing alga extracts with the alga organic fertilizer, freeze-drying, and spraying the obtained alga bioorganic fertilizer with chitosans and a sodium alginate solution. The organic fertilizer which contains various biological growth promoters can improve utilization rates of the nutrients by crops and increase endurances and validities of effective components, and also can increase ventilation and coalescence capabilities of soil, obviously promote the formation of granule structures of the soil, slowly release the nutrients, satisfy demands of plants of different growth stages, and generate obvious positive effects on the crop maturation, output and quality improvements, disease and insect pest resisting and the like.
Description
Technical field
The present invention relates to a kind of fertilizer and preparation method thereof, specifically, relate to a kind of chitosan coated seaweed bio fertilizer and preparation method thereof.
Background technology
Marine alga is to be grown in low in the ocean to wait spore plant, because its special growing environment, and very big different of nutritive ingredient and terrestrial plant existence; Contain incomparable abundant mineral element of general terrestrial plant and VITAMINs etc. in the marine alga; Seaweed fertilizer has been used as important liquid level spray in recent years, is applied to a lot of crops, comprises a lot of grass classes, cereal; Flowers and vegetable variety are all obtained effect preferably such as disease-resistant yield-increasing, synergy.Seaweed fertilizer derives from the synergy of 4 kinds of essential substance in the marine alga for the positively effect of crop yield and quality and health: carrageenin, plant hormone, amino acid and mineral substance, and carbohydrate.Carrageenin is as a kind of natural promoter, promotes the absorption of crop to other nutritive ingredients through absorption and osmosis, and amino acid and mineral substance can promote proteinic synthesizing and the stalwartness of plant tissue organ; Glucide in the marine alga is actually some polysaccharide (like Lalgine); Make a large amount of positively charged ion nutritive substances as a kind of natural sequestrant, also have iron to be absorbed by crop like copper, manganese, magnesium, calcium, polysaccharide is also as a kind of inductor; Stimulating growth is regulated the formation of material (polyamines) and natural antibiotics (plant antimycin); Through plant hormone and other compositions, such as the synergy of Sargassum polysaccharides, the ability that plant is resisted disease and pest has also obtained reinforcement; Promoted the healthy growth of crop; In addition, marine alga is to the also non-same common music of the improved effect of soil, and the seaweed fertilizer that contains materials such as Sargassum polysaccharides is natural soil redeposition.Use seaweed fertilizer in the soil, not only can chelating heavy metal-polluted soil ion, pollution abatement, and can promote the formation of soil aggregate also can directly or indirectly increase the soil organism.And in soil, Lalgine combines with metallic element, forms the cross-linked polymer of a kind of molecular weight multiplication, this polysalt in case and water molecules, just can control water molecules firmly.Compare with common fertilizer, seaweed fertilizer has more efficiently, is prone to absorb and eco-friendly characteristics.
About the research of seaweed fertilizer, mainly adopt the prepare seaweed fertilizer of seaweed extracted liquor in recent years, the mode of acquisition adopts the mode of chemical reagent lixiviate more, and the active substance loss is bigger.And very few with the research of microbial fermentation mode acquisition seaweed organic fertilizer, more do not see seaweed organic fertilizer is carried out the correlative study that coating is handled.
Summary of the invention
The purpose of this invention is to provide a kind of chitosan coated seaweed bio fertilizer and preparation method thereof.
In order to realize the object of the invention, a kind of chitosan coated seaweed bio fertilizer of the present invention, its preparation method is:
The extraction of A marine alga physiologically active substance: a) get after the marine alga removal of impurities cleans, squeeze the fritter that broken frond is 1-5cm, preferred 3cm; B) add entry and carry out secondary cover bubble and handle, c) with algae-residue with the acid solution processing that is hydrolyzed; D) with the centrifugal salinity that removes of hydrolyzed solution; E) will overlap the merging of bubble liquid and hydrolyzed solution, the concentrated marine alga physiologically active substance extracting solution that obtains; Wherein, Secondary cover bubble is handled and is specially: the water that in I pond and II pond, respectively adds 4～6 times of marine alga weight (preferred 5 times); Earlier first marine alga is put into the I pond and soak 50～80min (preferred 60min); Pull control out and change over to after clean and soak 50～80min (preferred 60min) in the II pond, the soaked in water in the I pond is poured out the back directly concentrate, in the I pond, add the water of marine alga weight 4～6 times (preferred 5 times) then again; After from the II pond, pulling marine alga out; Second batch of marine alga put into the II pond soak 50～80min (preferred 60min); Pull out to change over to behind the marine alga and soak 50～80min (preferred 60min) in the I pond; Soaked in water in the II pond is poured out the back concentrate, and in the II pond, add the water of 4～6 times of marine alga weight (preferred 5 times) again; After from the I pond, pulling marine alga out, the 3rd batch of marine alga put into the I pond soak 50～80min (preferred 60min), pull the marine alga continued out and change in the II pond and soak, so repeatedly, guarantee that promptly every batch of marine alga soaks twice, merge above-mentioned liquid concentrator and must overlap bubble liquid;
The preparation of B seaweed bio fertilizer: f) get the marine alga of proper amount of fresh and the algae-residue merging of steps A and mix thoroughly, stack retting rots, and smashes the back and adds chicken manure, stalk, Pericarppium arachidis hypogaeae, mixes back interpolation fermenting agent, carries out aerobic fermentation; G) add nitrogen phosphorus potassium, trace element and function yeast after the fermentation ends; H) with steps A in the seaweed extracted liquor that obtains merge lyophilize;
The preparation of the chitosan coated seaweed bio fertilizer of C: the seaweed bio fertilizer that obtains among the step B is sprayed chitosan and sodium alginate soln respectively, process chitosan coated seaweed bio fertilizer;
Wherein, said marine alga is one or more in sea-tangle, bulk kelp, bladder wrack, extra large capsule algae, Enteromorpha and the kelp;
Said fermenting agent is yeast, actinomycetes and bacterium;
Said acid solution is a hydrochloric acid soln, and the pH value of solution value was 2.5～3.0 when selective hydrolysis was handled;
Said function yeast is one or more in can subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus Licheniformis), bacillus megaterium (Bacillusmagaterium) and the jelly genus bacillus of commercially available acquisition.
The preparation method of chitosan coated seaweed bio fertilizer of the present invention comprises:
At first prepare seaweed extracted liquor, use fresh marine alga, remove impurity and exotic, rinse well, frond is squeezed, make frond broken; Add fresh water and overlap the bubble processing; Algae-residue is hydrolyzed with dilute acid soln; With hydrolyzed solution centrifugal remove behind the salinity subsequent use; To overlap bubble liquid and hydrolyzed solution and merge and concentratedly obtain marine alga physiologically active substance extracting solution, filter residue is subsequent use.
Next prepares the seaweed bio fertilizer, gets the marine alga of proper amount of fresh and algae-residue that a last step filters out and merges and mix thoroughly, and stack retting rots; Smash subsequently, evenly add chicken manure, stalk, Pericarppium arachidis hypogaeae, mix the back and add fermenting agent; Stir aerobe fermentation; Add nitrogen phosphorus potassium, trace element and function yeast, seaweed extracted liquor and seaweed bio fertilizer that a last step is obtained merge, and carry out drying.
The seaweed bio fertilizer that a last step is obtained places coater, sprays chitosan and sodium alginate soln respectively, processes chitosan coated seaweed bio fertilizer.
Wherein, said marine alga is one or more in sea-tangle, bulk kelp, bladder wrack, extra large capsule algae, Enteromorpha and the kelp;
Said fermenting agent is yeast, actinomycetes and bacterium;
Said acid solution is a hydrochloric acid soln, and the pH value of solution value was 2.5～3.0 when selective hydrolysis was handled;
Said function yeast is one or more in subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus Licheniformis), bacillus megaterium (Bacillus magaterium) and the jelly genus bacillus.
Advantage of the present invention and beneficial effect:
(1) contains several physiological active substances in the chitosan coated seaweed bio fertilizer provided by the invention, and the effect of slow-release or control-release fertilizer is arranged, have disease-resistant yield-increasing, many advantages such as native, nuisanceless of living.
(2) the present invention has adopted the broken method that combines with chemical treatment technology of efficient physical to extract marine alga marine alga physiologically active substance and nutritive substance; And adopted secondary cover bubble technology, at utmost kept long agent of each plant growth-promoting and multiple nutritional components in the bright marine alga.
(3) be added with bacteria group among the present invention, adopt the mode of aerobic fermentation to obtain seaweed organic fertilizer, and add nitrogen phosphorus potassium and an amount of trace element, comprehensive nutrition.
(4) the present invention adopts chitosan coated mode to prepare the seaweed bio fertilizer, and coated fertilizer is nontoxic, stable, inexpensive and the source is abundant, belong to environment-friendly material, and controlled-release effect is good.
(5) low in raw material cost of the present invention is easy to get, and preparation technology is simple, and production efficiency is high, and cost is low, and environmental pollution is few.
Description of drawings
Fig. 1 is the preparation flow figure of marine alga physiologically active substance extracting solution of the present invention.
Fig. 2 is the preparation flow figure of the chitosan coated seaweed bio fertilizer of the present invention.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.If do not specialize the conventional means that used technique means is well known to those skilled in the art among the embodiment, the raw materials used commercial goods that is.The percentage sign that relates among the embodiment " % " if do not specify, is meant mass percent.
Embodiment 1
At first be the preparation of marine alga physiologically active substance extracting solution, be specially: foreign material are removed in the sea-tangle flushing of gathering, be divided into many parts, every part of 1000kg; The squeezing machine squeezing is broken to be the fritter about 3cm, and I pond, II pond respectively add 5000kg fresh water, and (time is too short, is not enough to soak out active substance earlier sea-tangle to be soaked 60min in the I pond; Time is oversize, has increased the water-absorbent of sea-tangle, and then has increased the recovery of active substance, and the leaching yield of 60min can reach 85%; Can reach 94%), the soaked in water in I pond is leached concentrated, change the II pond over to after the sea-tangle control is clean, once more submergence 60min; Pull the fermentation that sea-tangle carries out next step out, the I pond adds new water, and the II pond replenishes sufficient fresh water, and the next batch sea-tangle is soaked 60min earlier in the II pond; Change I pond 60min again over to, so repeatedly, leach liquor concentrates; With the sea-tangle of soaking 60 ℃ of hydrolysis 6h under the condition of pH value 2.5 (adjusting) with hydrochloric acid; Centrifugal 5min under the hydrolyzed solution 4000rpm/min; Get supernatant and merge, till 50 ℃ of evaporation concentration to faint yellow sticky slightly shape, promptly obtain marine alga physiologically active substance extracting solution with cover bubble liquid.Preparation flow is as shown in Figure 1.
Next prepares the preparation of seaweed bio fertilizer; Specifically taking by weighing in fresh sea-tangle of 500kg and the above-mentioned steps filtering filter residue merges and mixes thoroughly; Stack retting rots, and smashes subsequently, evenly adds 100kg chicken manure, 100kg stalk, 100kg Pericarppium arachidis hypogaeae; Mix the back and add fermenting agent, contain bacterium, yeast and the actinomycetes (about 50 * 10 of balanced mix
8Individual viable bacteria/gram microbial inoculum), addition is 5% of a filter residue quality, stirs, and aerobic fermentation 40 days adds nitrogen phosphorus potassium, trace element and function yeast, and nitrogen phosphorus potassium addition is 5% of a filter residue quality; The trace element (boron, iron, copper) addition＞1%, and the addition of single-element must not＜0.2%; Function yeast is added Bacillus licheniformis (Bacillus Licheniformis) and subtilis (Bacillussubtilis); Addition is a 2kg/ ton filter residue; But the addition evenly distribute of two kinds of bacterium; But the living bacteria count of wanting the assurance function bacterium is not less than 200,000,000/g fertilizer, merges the seaweed extracted liquor in the last step and carries out drying.
The 3rd step was the preparation of chitosan coated seaweed bio fertilizer; Be specially and go up the seaweed bio fertilizer that obtains in the step, get in a certain amount of BY-300 of the being invested in type coater, 30 ℃ of the pot bodies of adjusting coater; Rotating speed 1300/rmin; The preheating several minutes sprays chitosan and sodium alginate soln respectively to the intravital seaweed bio fertilizer of pot then, sprays 5% chitosan (being prepared at 5: 1 by mass ratio by the chitosan of molecular weight 300,000 and the chitosan of molecular weight 2000) earlier; Spray 4% sodium alginate soln again; Notice that the used coating liquid quality of per unit weight must not be greater than 25%, whole process keeps in pot, passing to hot blast, to remove the moisture on seaweed bio fertilizer surface.Preparation flow is as shown in Figure 2.
Embodiment 2
At first be the preparation of marine alga physiologically active substance extracting solution, be specially: foreign material are removed in the Enteromorpha flushing of gathering, taken by weighing many parts, every part of 1000kg; The squeezing machine squeezing is broken to be the fritter about 3cm, and I pond, II pond respectively add 5000kg fresh water, earlier Enteromorpha is soaked 80min in the I pond; (according to the physical property of marine alga, but the soak time proper extension is to 80min) soak solution leaches concentrated, in the I pond, adds 5000kg fresh water again; Change the II pond over to after sea-tangle control is clean, submergence 80min once more, the II pond replenishes sufficient fresh water; The function in exchange I pond, II pond, so repeatedly, leach liquor concentrates subsequent use; With the Enteromorpha that soaked 60 ℃ of hydrolysis 6h under the condition of pH value 3.0 (adjusting) with hydrochloric acid; Centrifugal 5min under the hydrolyzed solution 4000rpm/min; Get supernatant and merge, till 50 ℃ of evaporation concentration to faint yellow sticky slightly shape, promptly obtain marine alga physiologically active substance extracting solution with cover bubble liquid.Preparation flow is as shown in Figure 1.
Next prepares the preparation of seaweed bio fertilizer; Be specially and get in fresh Enteromorpha of 500kg and the above-mentioned steps filtering filter residue and merge and mix thoroughly; Stack retting rots, and smashes subsequently, evenly adds 100kg chicken manure, 100kg stalk, 100kg Pericarppium arachidis hypogaeae; Mix the back and add fermenting agent, contain bacterium, yeast and the actinomycetes (about 50 * 10 of balanced mix
8Individual viable bacteria/gram microbial inoculum), addition is 5% of a filter residue quality, stirs, and aerobic fermentation 40 days adds nitrogen phosphorus potassium, trace element and function yeast, and nitrogen phosphorus potassium addition is 5% of a filter residue quality; The trace element (boron, iron, copper) addition＞1%, and the addition of single-element must not＜0.2%; Function yeast is added bacillus megaterium (Bacillus magaterium) and subtilis (Bacillussubtilis); Addition is a 2kg/ ton filter residue; But the addition evenly distribute of two kinds of bacterium; But the living bacteria count of wanting the assurance function bacterium is not less than 200,000,000/g fertilizer, merges the seaweed extracted liquor in the last step and carries out drying.
The 3rd step was the preparation of chitosan coated seaweed bio fertilizer; Be specially and go up the seaweed bio fertilizer that obtains in the step, get certain amount and be put in the BY-300 type coater, 30 ℃ of the pot bodies of adjusting coater; Rotating speed 1300/rmin; The preheating several minutes sprays chitosan and sodium alginate soln respectively to the intravital seaweed bio fertilizer of pot then, sprays 5% chitosan (being prepared at 5: 1 by mass ratio by the chitosan of molecular weight 300,000 and the chitosan of molecular weight 2000) earlier; Spray 4% sodium alginate soln again; Notice that the used coating liquid quality of per unit weight must not be greater than 25%, whole process keeps in pot, passing to hot blast, to remove the moisture on seaweed bio fertilizer surface.Preparation flow is as shown in Figure 2.
A kind of chitosan coated seaweed bio fertilizer provided by the invention and preparation method thereof.It has adopted the broken method that combines with chemical treatment technology of efficient physical to extract marine alga physiologically active substance and nutritive substance; And adopted secondary to overlap bubble technology; Make marine alga physiologically active substance extracting solution, can keep effective active composition and nutritive substance in the marine alga to greatest extent; Algae-residue is adopted special biochemical process, add multi-functional fermented bacterium, mainly contain yeast, actinomycetes and bacterium useful microbe, produce seaweed organic fertilizer with the mode of aerobic fermentation with synergism; Merge Seaweed Extract and seaweed organic fertilizer, drying; A certain amount of seaweed bio fertilizer is put in the coater, sprays chitosan and sodium alginate soln respectively, process chitosan coated seaweed bio fertilizer.These article can improve crop to reaching the utilization ratio of nutrient in the soil in the fertilizer; Increase the persistence and the validity of effective constituent; Also can increase the ventilative ability with coalescence of soil, obviously promote the formation of soil aggregate, and can slowly discharge nutrient; Satisfy the demand of different growth phase plants, aspects such as the raising of maturation, output and the quality of farm crop and opposing disease and pest have all been produced significant active effect.And being different from chemical fertilizer, seaweed organic fertilizer is easy to biological degradation, and is pollution-free, and the mankind, animal and birds are not had harm.
Application examples 1
With the 7000 mu of western muskmelons in somewhere is the planting base; Fertilizer is selected the chitosan coated seaweed bio fertilizer (experimental group) of preparation in the compound organic and inorganic fertilizer (control group) and the embodiment of the invention 1; The compound organic and inorganic fertilizer consumption is a 100kg/ mu, and the fertilizer consumption of embodiment 1 is a 70kg/ mu, and the result shows: experimental group is propped up 100 yuan than control group mu fertilising joint; More than the mu raising the output 200kg, the mu cutting down expenditures and increasing income is more than 1200 yuan.
Application examples 2
With 8 mu of orchards, somewhere is the apple cultivation base; Fertilizer is selected the chitosan coated seaweed bio fertilizer (experimental group) of preparation in the compound organic and inorganic fertilizer (control group) and the embodiment of the invention 2; The compound organic and inorganic fertilizer consumption is a 100kg/ mu; The fertilizer consumption of embodiment 2 is a 70kg/ mu, imposes the low-phosphorous high K composite fertilizer 30-40kg/ mu of high nitrogen 3-4 month in next year.The result shows: experimental group is propped up 150 yuan than control group mu fertilising joint, and the fruit tree growing way is good, and apple quality is good, and fruit-setting rate is high, and output on average increases by 20%, and the per mu yield value is crossed ten thousand yuan.
Though, the present invention has been done detailed description in the preceding text with general explanation and specific embodiments, on basis of the present invention, can to some modifications of do or improvement, this will be apparent to those skilled in the art.Therefore, these modifications or the improvement on the basis of not departing from spirit of the present invention, made all belong to the scope that requirement of the present invention is protected.
Claims (10)
1. chitosan coated seaweed bio fertilizer is characterized in that its preparation method is:
The extraction of A marine alga physiologically active substance: a) get after the marine alga removal of impurities cleans, squeeze broken frond; B) add entry and carry out secondary cover bubble and handle; C) with algae-residue with the acid solution processing that is hydrolyzed; D) with the centrifugal salinity that removes of hydrolyzed solution; E) will overlap the merging of bubble liquid and hydrolyzed solution, the concentrated marine alga physiologically active substance extracting solution that obtains;
The preparation of B seaweed bio fertilizer: f) algae-residue with fresh marine alga and steps A merges, and stack retting rots, and smashes the back and adds chicken manure, stalk, Pericarppium arachidis hypogaeae, mixes the back and adds fermenting agent, carries out aerobic fermentation; G) add nitrogen phosphorus potassium, trace element and function yeast after the fermentation ends; H) with steps A in the seaweed extracted liquor that obtains merge lyophilize;
The preparation of the chitosan coated seaweed bio fertilizer of C: the seaweed bio fertilizer that obtains among the step B is sprayed chitosan and sodium alginate soln respectively, process chitosan coated seaweed bio fertilizer;
Wherein, function yeast described in the step B is one or more in subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus Licheniformis), bacillus megaterium (Bacillusmagaterium) and the jelly genus bacillus.
2. fertilizer as claimed in claim 1; It is characterized in that; Secondary cover bubble is handled and is specially in the steps A: in I pond and II pond, respectively add the water of 4～6 times of marine alga weight, earlier first marine alga is put into the I pond and soak 50～80min, pull out to change in the II pond after the control only and soak 50～80min; Soaked in water in the I pond is poured out the back directly concentrate, in the I pond, add the water of 4～6 times of marine alga weight then again; After from the II pond, pulling marine alga out, second batch of marine alga put into the II pond soak 50～80min, pull out to change in the I pond behind the marine alga and soak 50～80min, the soaked in water in the II pond is poured out the back concentrate, and in the II pond, add the water of 4～6 times of marine alga weight again; After from the I pond, pulling marine alga out, the 3rd batch of marine alga put into the I pond soak 50～80min, pull the marine alga continued out and change in the II pond and soak, so repeatedly, guarantee that promptly every batch of marine alga soaks twice, merge above-mentioned liquid concentrator and must overlap bubble liquid.
3. according to claim 1 or claim 2 fertilizer is characterized in that said marine alga is one or more in sea-tangle, bulk kelp, bladder wrack, extra large capsule algae, Enteromorpha and the kelp.
4. according to claim 1 or claim 2 fertilizer is characterized in that acid solution described in the steps A is a hydrochloric acid soln.
5. according to claim 1 or claim 2 fertilizer is characterized in that fermenting agent described in the step B is yeast and bacterium.
6. fertilizer as claimed in claim 5 is characterized in that, fermenting agent described in the step B is actinomycetes.
7. the preparation method of each said chitosan coated seaweed bio fertilizer of claim 1～6 is characterized in that, comprises step:
The extraction of A marine alga physiologically active substance: a) get after the marine alga removal of impurities cleans, squeeze broken frond; B) add entry and carry out secondary cover bubble and handle; C) with algae-residue with the acid solution processing that is hydrolyzed; D) with the centrifugal salinity that removes of hydrolyzed solution; E) will overlap the merging of bubble liquid and hydrolyzed solution, the concentrated marine alga physiologically active substance extracting solution that obtains;
The preparation of B seaweed bio fertilizer: f) algae-residue with fresh marine alga and steps A merges, and stack retting rots, and smashes the back and adds chicken manure, stalk, Pericarppium arachidis hypogaeae, mixes the back and adds fermenting agent, carries out aerobic fermentation; G) add nitrogen phosphorus potassium, trace element and function yeast after the fermentation ends; H) with steps A in the seaweed extracted liquor that obtains merge lyophilize;
The preparation of the chitosan coated seaweed bio fertilizer of C: the seaweed bio fertilizer that obtains among the step B is placed coater, spray chitosan and sodium alginate soln respectively, process chitosan coated seaweed bio fertilizer;
Wherein, said function yeast is one or more in subtilis (Bacillus subtilis), Bacillus licheniformis (Bacillus Licheniformis), bacillus megaterium (Bacillus magaterium) and the jelly genus bacillus.
8. method as claimed in claim 7; It is characterized in that; Secondary cover bubble is handled and is specially in the steps A: in I pond and II pond, respectively add the water of 4～6 times of marine alga weight, earlier first marine alga is put into the I pond and soak 50～80min, pull out to change in the II pond after the control only and soak 50～80min; Soaked in water in the I pond is poured out the back directly concentrate, in the I pond, add the water of 4～6 times of marine alga weight then again; After from the II pond, pulling marine alga out, second batch of marine alga put into the II pond soak 50～80min, pull out to change in the I pond behind the marine alga and soak 50～80min, the soaked in water in the II pond is poured out the back concentrate, and in the II pond, add the water of 4～6 times of marine alga weight again; After from the I pond, pulling marine alga out, the 3rd batch of marine alga put into the I pond soak 50～80min, pull the marine alga continued out and change in the II pond and soak, so repeatedly, guarantee that promptly every batch of marine alga soaks twice, merge above-mentioned liquid concentrator and must overlap bubble liquid.
9. like claim 7 or 8 described methods, it is characterized in that acid solution described in the steps A is a hydrochloric acid soln.
10. like claim 7 or 8 described methods, it is characterized in that fermenting agent described in the step B is yeast and bacterium.
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