CN1772880A - Agricultural waste fermenting bacterial prepn and its prepn process - Google Patents

Agricultural waste fermenting bacterial prepn and its prepn process Download PDF

Info

Publication number
CN1772880A
CN1772880A CN 200510097902 CN200510097902A CN1772880A CN 1772880 A CN1772880 A CN 1772880A CN 200510097902 CN200510097902 CN 200510097902 CN 200510097902 A CN200510097902 A CN 200510097902A CN 1772880 A CN1772880 A CN 1772880A
Authority
CN
China
Prior art keywords
fermentation
microbial inoculum
substratum
compost
days
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN 200510097902
Other languages
Chinese (zh)
Other versions
CN100338211C (en
Inventor
薛智勇
王卫平
朱凤香
钱红
吴传珍
何福恒
章锦秋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang Academy of Agricultural Sciences
Original Assignee
Zhejiang Academy of Agricultural Sciences
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang Academy of Agricultural Sciences filed Critical Zhejiang Academy of Agricultural Sciences
Priority to CNB2005100979022A priority Critical patent/CN100338211C/en
Publication of CN1772880A publication Critical patent/CN1772880A/en
Application granted granted Critical
Publication of CN100338211C publication Critical patent/CN100338211C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Images

Abstract

The present invention discloses fermented bacteria preparation for agricultural waste treatment and its preparation process. The present invention separates and breeds three kinds of bacteria with fast growth and high heat resistance, including two strains of tectorial membrane yeast ZJY2 and Strreptomyces griseus W1 and one known strain Bacillus cereus ME1-2. These three kinds of spawn are cultured and fermentation produced separately and further produced into compound bacteria preparation in certain ratio or single bacteria preparation. When the bacteria preparation is used in treating agricultural waste, the waste is added with regulator to control the water content in 55-65 wt% and the bacteria preparation in 0.1-0.5 wt% is added through stirring, and the mixture is aerobic high temperature fermented for 20-30 days to reduce amount, reduce harm and become useful resource.

Description

Agricultural waste fermenting bacterial preparation and preparation method thereof
The application is original application day: on March 18th, 2004, and application number: 200410039818.0, invention and created name is divided an application for " being used for fermenting agent of agricultural wastes processing and preparation method thereof " application.
Technical field
The invention belongs to the fermenting agent and the manufacturing field thereof of handling husbandry and food-processing waste, especially belong to the microbial inoculum that to accelerate the material fermentation and the preparation method and the application thereof of this microbial inoculum.
Background technology
The livestock and poultry breeding scale industry of China was developed rapidly in the nearly more than ten years, and especially the pig industry that the intensive culture degree is very high has obtained significant progress, polluted but also caused than serious environmental when improving production level.In addition, produce every year such as a large amount of agricultural crop straw etc. in the agriculture production, and processing of farm products waste (as pericarp, leaf, slag and edible fungi residues etc.), more aggravated rural environmental pollution.These feces of livestock and poultry and agricultural wastes are source of pollution, it is again good organic fertilizer resource, through the microorganism aerobic thermophilic fermentation, realizing minimizing (material moisture, volume, weight reduce), innoxious (eliminate objectionable impurities, kill parasite, ovum, pathogenic micro-organism, grass-seed etc.) processing and resource utilization (organic fertilizer, nutraceutical matrix etc.) utilization, is one of effective way of alleviating agricultural pollution and Sustainable development husbandry.The farmland is abandoned or applied to the most of organic waste in current rural area unprocessed just directly discharging, heap, has small part then to adopt the way of traditional composting spontaneous fermentation.But the organism of fermentation flora starting quantity of natural compost is few, propagation is slow, and non-refractory often, so that compost fermentation heats up is slow, fermentation period is long, needs more than 2 months usually.Because fermentation heap Wen Bugao, it is insufficient to ferment, and harmful organism is killed not thorough.
Summary of the invention
The objective of the invention is,, go out to have the microorganism strains of characteristic such as high temperature resistant, energetic, that growth is fast from the directed isolation and selection of nature at above deficiency; After cultivating propagation, make and provide fermentation microbial inoculum product and preparation method thereof; The technology that also provides simultaneously this microbial inoculum acceleration husbandry of application and food-processing fermentation of organic wastes to become thoroughly decomposed.
Purpose of the present invention is achieved by the following technical programs.
The microorganism of using
Microbial inoculum of the present invention is through cultivation, propagation, formulated by following three kinds of bacterial classifications.Bacterial strain uses therefor is saccharomycopsis fibuligera (Saccharomycopsis fibuligera) ZJY 2, grey gamboge streptomycete (Streptomyces griseoluteus) W1 and bacillus cereus (Bacillus cereus) ME1-2.In 1997 from Hangzhou, different areas such as Wenzhou and Beijing, Shenyang, Nanjing gather high temperature compost and pedotheques such as cow dung, pig manure, chicken manure and rubbish, utilize the conventional substratum of inhomogeneity bacterial classification therefrom orientation isolate totally 71 strains of yeast, actinomycetes and genus bacillus, and carry out the bacterial strain purifying, and then bacterial strain is shaken bottle by 44 ℃ of inclined-planes and 50 ℃ carry out high temperature resistant cultivation and mensuration, it is as follows therefrom to filter out high-temperature resistant strain:
Saccharomycopsis fibuligera ZJY 2(be designated hereinafter simply as ZJY 2Bacterial strain), this bacterial strain is in March, 1997, separates and gets from Zhejiang Hangzhou with aforesaid method.Through the following experimental result of Institute of Microorganism, Academia Sinica's foundation, the key analysis of contrast " YEASTS:Characteridtics and Identification " determines that this yeast is the saccharomycopsis fibuligera (Saccharomycopsisfibuligera) in the Saccharomycopsis (Saccharomycopsis).Its main difference point is that this strain growth speed is fast, easily cultivates, and is difficult for polluting, and the substratum preparation is simple, anti-high temperature (40 ℃ of oven dry are not dead) to a certain degree.This new bacterial strain on March 24th, 2003 in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCCNO.0909.
Ash gamboge streptomycete W1 (being designated hereinafter simply as the W1 bacterial strain), this bacterial strain is in March, 1997, separates from Wenzhou, Zhejiang and gets with aforesaid method.Through Institute of Microorganism, Academia Sinica, by " actinomycetic classification and evaluation ", " streptomycete identification handbook ", " actinomycetes research and application ", in conjunction with following morphological specificity, cell walls chemical composition and cultural characteristic and every physiological and biochemical property, W1 bacterial strain and grey gamboge streptomycete are quite similar, are grey gamboge streptomycete (Streptomyces griseoluteus) so the W1 bacterial strain is named.Its main difference point is that this strain growth speed is fast, and increment is big, and high temperature resistant (50 ℃, 5 hours are not dead, grow fast when temperature is suitable) are easily cultivated.This new bacterial strain on January 9th, 2004 in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCC NO.1086.
Bacillus cereus ME1-2, this bacterium separated and gets from ShaoXing,ZheJiang with aforesaid method in May, 1989, identify through Zhejiang Agricultural Univ, according to utilization of carbon source, features such as enzyme reaction, by " classification such as outstanding Bacteria Identification handbook of uncle and R.E.Gordon determine that this bacterial strain belongs to the bacillus cereus of bacillus (Bacillus cereus), are known bacterial classification.The report document is published in " soil and fertilizer " 1993 the 2nd phases, 29~32 pages, " development of multiple-effect bacterium strain improvement, evaluation and microbial inoculum ", author He Fuheng, Zhang Jinqiu, Xue Zhiyong etc., " Zhejiang agricultural journal " the 5th the 2nd phase of volume in 1993,89~93 pages, " main characteristic of multiple-effect bacterium and preparation thereof ", author He Fuheng, Zhang Jinqiu, Xue Zhiyong etc.The public can obtain this microbial strains from the possessor (patent inventor) of this microbial strains.
ZJY 2Bacterial strain, W1 bacterial strain and ME1-2 have following character
1.ZJY 2Bacterial strain:
Colony characteristics, cellular form and physico-chemical property: on wort and YEPD substratum plate, 28 ℃ of static cultivations of constant temperature were observed in 2~4 days, and the bacterium colony surface is secretly-canescence, center projections, surface irregularity.On malt juice liquid medium, 24 hours microscopies of 28 ℃ of shaking culture are observed: modes of reproduction is bilateral or polygon budding.Cell is avette, oval and cured microscler, and individual cells is taper.The cell size is (2.0-2.5) * (3.3-12.5) μ m.On inoculation YEPD, the wort agar substratum, cultivate after 3 days for 28 ℃ and produce thecaspore, ascus sphere-avette.Each ascus contains 1~4 thecaspore.The weak glucose fermentation of this bacterium energy, sucrose, maltose, monose and disaccharide such as unfermentable semi-lactosi, lactose, raffinose, trehalose.Can assimilate glucose, sucrose, maltose, trehalose, raffinose, cellobiose, dextrin, Zulkovsky starch, ethanol, moss sugar alcohol, glycerine, SODIUMNITRATE, succsinic acid, saligenin etc.Can not assimilate lactose, close disaccharides, synanthrin, wood sugar, D-pectinose, ribose, rhamnosyl, glucosamine, methyl alcohol, N-acetylglucosamine, galactitol etc.In addition also can normal growth in the substratum that contains 10%NaCl and 5% glucose.Generally 37 ℃ of growths.Measure through high thermal resistance, better 45 ℃ of incubation growth.
2.W1 bacterial strain:
Colony characteristics, cellular form and physico-chemical property: 28 ℃ of inserted sheets were cultivated 7~10 days on Gao Shi synthetic No. one and sucrose Czapek's agar, and microscopy is observed and shown that the substrate mycelium of bacterial strain does not have tabula, does not rupture; The aerial hyphae bulk-growth is luxuriant, multi-branched; The straight shape of fibrillae of spores, what have has a gentle Qu Chengcong shape; Spore is more than 10.Spore oval or short cylinder, smooth surface.
The cultural characteristic of W1 bacterial strain on 7 kinds of substratum
Substratum Aerial hyphae Substrate mycelium Soluble pigment
The synthetic agar inorganic salts starch agar oatmeal agar potato of the synthetic agar sucrose Czapek's agar glycerine asparagine agar Ke Shi of Gao Shi is soaked juice agar The light grey dirt grey of the little long canescence snow weasel grey of gas silk pearl oyster grey dark grey look grey at night The yellow shallow gamboge look yellowish-brown yellowish-brown gamboge look grass brown of the yellow butterfat of almond Do not have
Analyze through the cell walls chemical composition, the strain whole-cell hydrolyzed solution contains LL-DAP (left-handed diaminopimelic acid, Diaminopimelic acid), glycine; Atypism sugar (sugared type C).The cell walls chemical composition belongs to the I type.Energy glucose fermentation, pectinose, fructose, N.F,USP MANNITOL, semi-lactosi, rhamnosyl, nonfermented raffinose, wood sugar, inositol, liquefy gelatin, milk is peptonized, but do not solidify milk, the starch hydrolysis positive, can reduce nitrate, not grow on the Mierocrystalline cellulose, not produce H 2S and class melanochrome.Measure through high thermal resistance, well-grown in 50 ℃ of high temperature, the mycelium pellet dry weight reaches 2.17g/60ml.
3. bacillus cereus ME1-2:
Colony characteristics, cellular form and physico-chemical property: colony characteristics is a subcircular, open and flat, and the edge is like the fringing shape, and is opaque, the wax shape; Cellular form is a Gram-positive, direct rod shape, and the cell size is (1.2~1.5) * (2.0~3.5) μ m.The dyeing of children's bacterium carbolfuchsin in age, cell has non-staining bead.Gemma is oval, and sporangiocyst does not expand, motion whole body flagellum.Energy glucose fermentation, wood sugar, trehalose, maltose, unfermentable pectinose, N.F,USP MANNITOL, lactose can be grown in 7% sodium-chlor nutrient solution, can reduce nitrate, have ammonification, can utilize Citrate trianion, indole test, VP react negative, urase, catalase, lecithinase are determined as the positive, do not utilize malonate, the energy hydrolyzed starch, on the nutrient broth of PH5.7, grow, hydrocellulose is not obvious, energy anaerobic growth, the negative reaction of tyrosine hydrolysis.Measure through high thermal resistance, better 45 ℃ of incubation growth.
Strain culturing and fermentation
Saccharomycopsis fibuligera ZJY2 is that the basis adds small amounts of inorganic salt such as calcium, magnesium, phosphorus etc. and carries out liquid shaking bottle fermentation, 25~35 ℃ of cultivations, rotating speed 120~220rpm with the malt extract medium.Make the female kind of solid as bacterial classification inoculation solid fermentation koji tray, Gu bent raw material is a wheat bran, 0~30%) and small amounts of inorganic salt etc. dish cake and soybean cake powder (ratio 100%~40%: 0~30%:, water content is 50~65%, cultivated 16-26 hour for 25-35 ℃, be inoculated in the wheat bran of sterilization again by 0.5~5% inoculum size, the dish cake, rice bran and soybean cake powder (ratio 100%~10%: 0~30%: 0~30%: 0~30%) and in the small amounts of inorganic salt mixed fermentation solid medium, water content is 40~65%, mix thoroughly and stack 30min after 0.01MPa sterilization 60min, 16-26 hour ventilation koji tray fermentation or Quchi or cultivate batch production production with solid-state fermentation tank, temperature is controlled at 20~38 ℃, the zymophyte number gets final product more than reaching 100,000,000 cfu/ gram, 40~50 ℃ of dryings are pulverized.
Ash gamboge streptomycete W1 be that basic medium adds small amounts of inorganic salt such as calcium, magnesium, phosphorus etc. and carries out liquid shaking bottle and ferment with Zulkovsky starch 2~6%, yeast powder 0.1~1.0%, soybean cake powder 5~30% etc., pH is adjusted into 5~9, rotating speed 120~220rpm cultivated 36~48 hours for 20~40 ℃.The inoculum size of planting by 1~10% as mother is inoculated in the solid fermentation substratum of sterilization, 25~10%) and small amounts of inorganic salt such as calcium, magnesium, phosphorus, potassium etc. solid fermentation substratum main component is wheat bran, soybean cake powder, a rice chaff (ratio 25~70%: 50~20%:, moisture content in medium is 50~65%, temperature is controlled at 35~37 ℃, aerated culture 48~72 hours, the zymophyte number gets final product more than reaching 0.5 hundred million/gram, and 40~50 ℃ of dryings are pulverized.
Bacillus cereus ME1-2 shakes bottle with the inoculation of beef extract-peptone slant strains earlier and makes liquid seeds, the liquid shaking bottle substratum is that basic medium adds small amounts of inorganic salt such as calcium, magnesium, phosphorus, potassium etc. and is prepared from the beef extract-peptone, adjust pH 7.0~7.8, rotating speed 120~220rpm cultivated 8 to 16 hours for 28 ℃~35 ℃.The inoculum size of planting by 5~10% as mother is inoculated in liquid fermentation tank cultivation batch production production, the liquid fermentation tank substratum is Semen Maydis powder 0.1~0.6%, yeast powder 0.1~0.5%, soybean cake powder 0.1~0.6%, glucose or sucrose 0.1~0.3% and small amounts of inorganic salt are formed, and surplus is a water, and temperature is controlled at 25~35 ℃, aerated culture 18-30 hour, the zymophyte number gets final product more than reaching 100,000,000 cfu/ gram, adsorbs mixing by 1: 1 (v/w) with wheat bran, 40~45 ℃ of dryings are pulverized.
The preparation of microbial inoculum
Above-mentioned three kinds of bacterium are single microbial inoculum of planting after cultivation and fermentation is pulverized respectively, three kinds or wantonly two kinds of parts by weight ratios in 10~1: 0~3: 0~6 are carried out the compound composite fungus agent that is, can find out pig manure fermentation test result according to different microbial inoculum combinations, with ZJY2, three kinds of microbial inoculums of W1 and bacillus cereus ME1-2 carry out compound in 4: 3: 3 parts by weight ratio, this preferred proportion compound microbial inoculum is to pig manure leavening temperature and the best (see figure 1) of dehydrating effect.Above-mentioned composite fungus agent or single bacterium number average of planting microbial inoculum should be controlled at more than the 0.5 hundred million cfu/ gram, and water ratio should be controlled between 8~14%.The packing solid powder can be with moistureproof woven bag or plastics bag, and liquid bacterial agent can be with sealed vessel or packed.
Microbial inoculum of the present invention was at room temperature stored 6 months through test, and the bacterium number is 12.17 * 10 8Cfu/g, survival rate is up to 84.11%; The bacterium number of storing 9 months is 9.53 * 10 8Cfu/g, survival rate 65.86%; The bacterium number of storing after 1 year still reaches 2.27 * 10 8Cfu/g, survival rate is 18.65%.
Microbial inoculum of the present invention is checked according to " toxicological evaluation of food safety procedure " through Zhejiang Academy of Medical Sciences province laboratory animal and safety research key lab, microbial inoculum is fed maximum tolerated dose greater than 40.0g/kg to the ICR its mouse oral, experimental session, mouse non-toxic reaction and death, and weight increase, true border is nontoxic.The ME1-2 microbial inoculum adopts " karber's method " per os to irritate stomach acute toxicity test, female, male mouse LD to laboratory animal NIH mouse (female, male grouping) 50>20g/kg belongs to nontoxic substantially.So microbial inoculum of the present invention might be used as food or fodder additives.
Experimental record to its mouse oral administration maximum tolerated dose
Group Dosage (g/kg) Number of animals (only) Sex Toxic reaction Body weight (g, X ± SD)
Before the administration Five days Ten days
ZJY2 40.0 10 No abnormal 22.5±0.97 26.5±1.27 27.7±1.64
No abnormal 22.5±0.97 29.3±1.57 31.4±1.90
W1 40.0 10 No abnormal 22.6±0.84 27.0±1.56 28.4±1.78
No abnormal 22.5±0.97 29.4±1.43 31.0±2.11
The application of microbial inoculum in compost fermentation
According to China, especially the scale raising of southern area pig is maximum, number of animals raised maximum, and pig manure water ratio height (70~90%), sticky, ventilation property is poor, and C/N is than low, the characteristics of spontaneous fermentation difficulty, microbial inoculum of the present invention have proposed concrete application method based on the fresh pig dung fermentation test.
(1) by adding amendment, as wood chip, rice chaff, Chinese medicine slag, mushroom slag etc., from 70~90% falling tones to 55~65%, addition is generally about 5~30% of feces of livestock and poultry the like waste total amount the water ratio of pig manure, and preferred addition is 10~15%;
(2) the microbial inoculum inoculum size is preferably 2 ‰ with heavy 1 ‰~5 ‰ being advisable of fermentation materials (being the mixture after waste adds amendment); Turn evenly after bacterial classification adds, the compost height generally is advisable with 80~100 centimeters, is beneficial to aerobic fermentation and moisture evaporation;
(3) turning regularly, ventilation; Left and right sides turning in 1~3 day once, both ventilates promotes compost fermentation, especially can obviously improve compost temperature in early stage and rise; Quicken the compost moisture evaporation again, reach minimizing; And the compost core temperature is descended, the normal bacterial classification that makes peripheral shallow-layer is accelerated the fermentation of new round aerobic high-heat along with material enters internal layer, thereby accelerates the fermentation maturity process of whole compost.
(4) become thoroughly decomposed standard and quality examination of compost fermentation
Compost continuously fermented more than 55~60 15~20 days the time, soaked 1 day by 1: 10 with tunning and clear water, observe vat liquor and be coffee-like, piling temperature simultaneously begins to descend, show that then the biological fermentation material enters the stage of becoming thoroughly decomposed, the continuity fermentation is after 3~5 days, through drying or dry pulverizing again, water content is controlled at below 20%, organic fertilizer product.Require coliform count in this product<10 5Individual/kg, roundworm egg is counted mortality ratio>95%, reaches innoxious requirement (livestock and poultry breeding industry pollutant emission standard GB18596-2001), and quality product meets national agricultural industry criteria (NY525-2002).
All the other feces of livestock and poultry and agricultural organic waste except that pig manure should be according to its water content height, air permeability quality, above-mentioned way is done suitably to adjust, be lower than 60% as the fermentation materials water ratio, then do not need to add amendment etc., all can obtain fermentation maturity effect preferably.
Beneficial effect of the present invention: after one, microbial inoculum of the present invention is applied to feces of livestock and poultry and agricultural organic waste compost treatment, it is fast that show fermentation calefaction early stage, can reach more than 55~65 ℃ in initial 5 days, 10 days temperature of compost are near 70 ℃ of peak values, and the compost that 15 days internal ratios do not connect bacterium improves 5~11 ℃; Middle and later periods can keep higher leavening temperature, medial temperature contrast (spontaneous fermentation) is high 8~10 ℃, thereby accelerated whole fermentation maturity process, the fermentation maturity phase shortens to 20 days to 30 days by contrast more than 2 months, improve processing place usage ratio of equipment greatly, reduced production cost.The influence that the fermentation heap material heats up, the fermentation maturity process is subjected to climatic factor, the cycle in summer is shorter, and prolong winter.Two, because fermentation heats soon, the holding temperature height can make coliform count in the compost and roundworm egg number etc. reach innoxious requirement, weed seed death in general about 10 days.Three, composting material leavening temperature height has also been accelerated the evaporation of fermentation materials moisture.In the general fermentation 20~30 days, can make the initial water ratio of material descend about 30%, reach the minimizing requirement, also obtain deodorizing effect preferably simultaneously.Four, the bacteria fermentation fertilizer is safe in utilization respond well.The present invention adopts the germination index test of cucumber seeds to show that pig manure fermented to 20~30 days, and its germination index (GI%) is worth all more than 70%, shows plant safety; Fertilising is after discharge to measure soil nutrient, show comparatively steadily, and nutrient preserving capability is good, and can activating rhizosphere soil with increase soil fertility, bacterium, actinomycetes, genus bacillus, fungi etc. all roll up in the soil, have improved edatope and crumb structure; Clear and definite through the crop application test, this fertilizer increases vitamins C, sugar equal size to output that is improved such as vegetables, melon, sugarcane, citrus, strawberries, and reduces the nitrate content of gourd, fruit and vegetable edible portion, is beneficial to HUMAN HEALTH.Five, better economic and social benefit are arranged, through measuring and calculating, about 250 yuan/ton of this fermentation organic fertilizer production and cost of goods sold, market price can reach about 350 yuan/ton.Therefore, the present invention can be turned waste into wealth, and is applicable to the fermentative processing of mass-producing livestock and poultry pasture ight soil and agricultural organic waste, and can form the industrialization development of fermenting agent and organic commercial fertilizer, improves agriculture benefit.
Description of drawings
The combination of Fig. 1 different strain is to the synoptic diagram that influences of pig manure leavening temperature and water ratio
Fig. 2 inoculates the influence synoptic diagram of microbial inoculum to compost temperature and germination index
Fig. 3 microbial inoculum different vaccination amount influences synoptic diagram to pig manure leavening temperature and dehydrating effect
Embodiment
By being described in more detail the present invention by following examples.Following examples only are illustrative, and the present invention is not subjected to the restriction of these embodiment.
One, pulvis Production Example:
1, the preparation of saccharomycopsis fibuligera bacterial strain ZJY2 microbial inoculum:
Saccharomycopsis fibuligera ZJY2 liquid shaking bottle substratum: wort (10 ° of Bx) 1000ml, glucose 18 grams, pH nature.Promptly be prepared into the female inclined-plane of planting of solid as substratum adding 20 gram agar and carry out inoculation culture, get the female ring of planting in inclined-plane by the aseptic technique method and be inoculated in yeast liquid shaking bottle (the dress 60ml substratum/250ml triangular flask) substratum, 28 ℃, rotating speed 180rpm, cultivated 18 hours, the bacterium number reaches 0.5 * 10 8Cfu/ml is above to get final product.Saccharomycopsis fibuligera solid fermentation koji tray substratum: with wheat bran 99% and small amounts of inorganic salt MgSO 4, NaCl each 0.05%, by material, water adds water than 1: 1 (w/w) makes material moisture be about 60%.Liquid bacterial agent with shake-flask culture is a bacterial classification, the inoculum size by 0.1% evenly be inoculated in the solid fermentation material song (koji tray is wooden, and the length and width height is respectively 60cm, 45cm, 6cm, 3~5cm), 25 ℃ of cultivations of solid medium thickness, 20 hours, the bacterium number reached 2 * 10 8Cfu/g gets final product, and 40~50 ℃ of dryings are pulverized and made the female kind of solid.If adopt mass-producing koji tray fermentation or Quchi or solid-state fermentation tank production; its substratum is with solid fermentation koji tray substratum; after substratum is mixed stacking 30min thoroughly by material-water ratio; earlier through 0.1MPa sterilization 60min; inoculum size by 3% is inoculated in the koji tray solid fermentation substratum; temperature is controlled at 26 ℃; 26 hours koji tray aerobic fermentations or Quchi or cultivate batch production production with solid-state fermentation tank; the zymophyte number gets final product more than reaching 100,000,000 cfu/ gram; 40~50 ℃ of dryings; pulverized 40 mesh sieves, water content about 14% is this bacteria powder product.
2, the preparation of grey gamboge streptomycete W1 microbial inoculum:
Ash gamboge streptomycete W1 liquid shaking bottle substratum: Zulkovsky starch 2.5%, yeast powder 0.3%, soybean cake powder 5% are basic medium, add NaCl 2%, KH 2PO 40.05%, CaCO 30.4%, pH is adjusted into 7.2.Earlier adding 2% agar as substratum promptly is prepared into the female inclined-plane of planting of solid and carries out inoculation culture, getting female kind the in inclined-plane by the aseptic technique method produces grey gamboge streptomycete bacteria suspension 1ml with sterilized water and is inoculated in yeast liquid shaking bottle (the dress 100ml substratum/500ml triangular flask) substratum, rotating speed 160rpm, cultivated 48 hours for 20~40 ℃, bacteria fermentation biomass (dry weight) reaches 0.5g/100ml and gets final product.Each raw material weight per-cent of ash gamboge streptomycete solid fermentation substratum is: wheat bran 68.06%, soybean cake powder 20%, rice chaff 10% and NaCl 1.5%, KH 2PO 40.04%, CaCO 30.4%; add water in addition again; making moisture content in medium is 55%, and cultivating microbial inoculum with liquid shaking bottle serves as that female inoculum size of planting by 1% is inoculated in the solid fermentation substratum of sterilization, and temperature is controlled at 32 ℃; aerated culture 72 hours; the zymophyte number gets final product more than reaching 0.5 hundred million/gram, and grey gamboge streptomycete mass-producing solid fermentation method is the same, 50 ℃ of dryings; pulverized 40 mesh sieves, water content about 14% is this bacteria powder product.
3, the preparation of bacillus cereus ME1-2 microbial inoculum:
Bacillus cereus ME1-2 liquid shaking bottle substratum: select beef-protein medium for use, adjust pH 7.2.Be inoculated in liquid shaking bottle (the dress 60ml substratum/250ml triangular flask) substratum with beef extract-peptone slant strains one ring earlier by the aseptic technique method, 28 ℃, rotating speed 200rpm cultivated 16 hours, and the zymophyte number promptly can be used as liquid seeds more than reaching 100,000,000 cfu/ gram.Liquid fermentation tank substratum: Semen Maydis powder 1%, yeast powder 1%, soybean cake powder 1%, dextrin 1%, MgSO 47H 2O 0.07%, KH 2PO 40.1%, CaCO 30.5%, rape oil 0.3%, and pH is adjusted to 7.4~7.4 before the sterilization, sterilizes 2 hours for 121 ℃.Cultivating microbial inoculum with liquid shaking bottle serves as that female inoculum size of planting by 5% is inoculated in 6 tons of liquid fermentation tanks cultivations, temperature is controlled at 28~30 ℃, aerated culture 26 hours, batch production is produced, the zymophyte number gets final product more than reaching 1,000,000,000 cfu/ gram, adsorbs 50 ℃ of dryings by 1: 1 (v/w) with wheat bran, pulverized 40 mesh sieves, water content about 14% is this bacteria powder product.
4, the preparation of composite fungus agent:
Above-mentioned three kinds of microbial inoculums were in 4: 3: 3 (w/w/w) ratio uniform mixing, and microbial inoculum quantity reaches more than 0.5 hundred million/gram, and packing is three bacterial classification composite fungus agents.
Adopt above-mentioned preparation saccharomycopsis fibuligera ZJY2 microbial inoculum and bacillus cereus ME1-2 microbial inoculum or grey gamboge streptomycete W1 microbial inoculum, two kinds of microbial inoculums were in 7: 3 (w/w) ratio uniform mixing, microbial inoculum quantity reaches 0.5 hundred million/more than the gram, two bacterial classification composite fungus agents, packing gets final product.
Two, liquor Production Example
1, the preparation of saccharomycopsis fibuligera bacterial strain ZJY2 microbial inoculum:
Saccharomycopsis fibuligera ZJY2 liquid nutrient medium: wort (10 ° of Bx) 1000ml, glucose 18 grams, pH nature.Earlier adding 2% agar as substratum promptly is prepared into the female inclined-plane of planting of solid and carries out inoculation culture, getting the female ring of planting in inclined-plane by the aseptic technique method is inoculated in yeast liquid shaking bottle (the dress 60ml substratum/250ml triangular flask) substratum, 28 ℃, rotating speed 180rpm, cultivated 18 hours, the bacterium number reaches 0.5 * 10 8More than the cfu/ml, pack by aseptic technique and to be this bacterium liquor product.
2, the preparation of grey gamboge streptomycete W1 microbial inoculum
Ash gamboge streptomycete W1 liquid nutrient medium: Zulkovsky starch 2.5%, yeast powder 0.3%, soybean cake powder 5% are basic medium, add NaCl 2%, KH 2PO 40.05%, CaCO 30.4%, pH is adjusted into 7.2.Earlier adding 2% agar as substratum promptly is prepared into the female inclined-plane of planting of solid and carries out inoculation culture, getting female kind the in inclined-plane by the aseptic technique method produces grey gamboge streptomycete bacteria suspension 1ml with sterilized water and is inoculated in yeast liquid shaking bottle (the dress 100ml substratum/500ml triangular flask) substratum, rotating speed 160rpm, cultivated 48 hours for 20~40 ℃, bacteria fermentation biomass (dry weight) reaches 0.5g/100ml, packs by aseptic technique and is this bacterium liquor product.
3, the preparation of bacillus cereus ME1-2 microbial inoculum
Press bacillus cereus ME1-2 microbial inoculum pulvis method and produce, reach more than 1,000,000,000 cfu/ml,, directly pack and be this bacterium liquor product by aseptic technique without wheat bran absorption at the zymophyte number.
4, the preparation of composite fungus agent
By aseptic technique method uniform mixing, microbial inoculum quantity reached more than 0.5 hundred million/ml above-mentioned three kinds of microbial inoculums, packs to be three bacterial classification composite fungus agent liquor products in 4: 3: 3 (V/V/V) ratios.
Adopt above-mentioned preparation saccharomycopsis fibuligera ZJY2 microbial inoculum and bacillus cereus ME1-2 microbial inoculum or grey gamboge streptomycete W1 microbial inoculum, two kinds of microbial inoculums were in 7: 3 (w/w) ratio aseptic technique method uniform mixing, microbial inoculum quantity reaches more than 1,000,000,000 cfu/ml, pack two bacterial classification composite fungus agent liquor products.
Three, experimental example:
Microbial inoculum of the present invention and Japanese enzymatic microorganism, Beijing " organic waste zymophyte song ", do not inoculate contrast, be that raw material adds 15% rice chaff and an amount of microbial inoculum and carries out the compost fermentation simultaneous test with the fresh pig dung, the results are shown in following table, test site is in the Xiaoshan, Zhejiang city, in June, 2003 time, enzymatic microorganism is purchased the green precious ferment biological engineering corporation in Zhejiang, and " organic waste zymophyte song " purchased in Beijing Jingpuyuan Bio-engineering Co., Ltd..
Handle Microbial inoculum price (unit/ton) Microbial inoculum addition (‰) The fermentation fate (my god) Water ratio reduces (%) Heap temperature after 5 days (℃) Innoxious index Mass-producing, standardized production difficulty or ease
The bent microbial inoculum of the present invention of contrast (nature) ferment bacterium debirs zymophyte 0 8000 30000 5000 0 3 3 2 >30 30 25 20 <30 >30 >30 >30 39 55 56 60 Difficult up to standard up to standard easily up to standard Difficulty easily easily
Four, application examples:
1, animal waste is handled
Fresh pig dung (water ratio about 70%) adds amendment rice chaff 15%, the inoculum size inoculation fermenting agent of the present invention (three bacterium mix bacterium agents) by 3 ‰, turn over even after, pile high 80 centimetres, not inoculate microbial inoculum in contrast.The initial water content of fermentation muck is 62.38%.Turn over once heap every 3 days, put down in writing the germination index of the gentle 5 days sampling and measuring compost of heap of heat zone every day.The result shows (see figure 2), compost is from germination index (GI%) measurement result, the inoculation fermentation microbial inoculum compost in the time of 15 days near 70%, and do not connect bacterium will compost in the time of 25 days just near 70%, 5 days heap temperature of the compost of inoculation fermentation microbial inoculum has reached 61 ℃, than not increased by 8 ℃ and 36 ℃ with microbial inoculum and initial compost temperature, in preceding 15 days of compost fermentation, the compost fermentation temperature that the ratio of inoculation or not improves 8~11 ℃ respectively; The compost dewatering speed is also very fast, the (see figure 3) comparison is according to reducing by 11 percentage points, measure through agricultural product quality inspection center of the Ministry of Agriculture through the organic fertilizer product that the fermenting agent inoculation fermentation is obtained, quality product, coliform count, induced worm egg death rate all meet country and industry standard.Waiting under the cost situation, using the biological fermentation fertilizer in the fresh edible maize field than increasing production 5.7% with the dish cake; Comparison is according to volume increase 10.8%, and volume variance reaches utmost point conspicuous level; The bright preceding high-quality tea of using the biological fermentation fertilizer increases income 140 yuan than dish cake volume increase 18.5%, and common spring tea volume increase 24.2% increases income 76.5 yuan, adds up to increase income 216.5 yuan, and difference reaches utmost point conspicuous level.
2, fermenting crops straws is handled
Agricultural crop straw (or vegetables processing waste) dried be crushed to 3-5cm, add 20% fecaluria waste water, 0.3% bicarbonate of ammonia, inoculum size inoculation fermenting agent of the present invention (three bacterium mix bacterium agents) mixing by 3 ‰, pile high 100 centimetres, not inoculate microbial inoculum in contrast.The initial water content of fermented stalk material is 60.2%, turned over once heap every 3 days, put down in writing the heap temperature of heat zone every day, 5 days heap temperature of the compost of inoculation fermentation microbial inoculum has reached 58 ℃, respectively than not increased by 6 ℃ and 13 ℃ with microbial inoculum and initial compost temperature, in preceding 12 days of compost fermentation, the compost fermentation temperature that the ratio of inoculation or not improves 9 ℃; The compost dewatering speed is also very fast, and comparison is according to reducing by 11 percentage points, the organic fertilizer product that is obtained through the fermenting agent inoculation fermentation after measured, quality product meets industry standard.Waiting under the cost situation, using the biological fermentation fertilizer, on the vegetables tomato, using the biological fermentation fertilizer than increasing production 5.1% with the dish cake; Comparison is according to volume increase 8.7%, and every mu increases income 124.5 yuan.
3, be used for cultivation matrix production
Fresh pig dung (water ratio about 70%) 40%, rice chaff 20%, mushroom slag 10%, mushroom mud 20%, Chinese medicine slag 10%,, inoculum size inoculation fermenting agent of the present invention (three bacterium mix bacterium agents) by 3 ‰, turn over even after, pile high 80 centimetres, not inoculate microbial inoculum in contrast.The initial water content of fermented substrate is 55.6%.Turn over once heap every 3 days, put down in writing the germination index of the gentle 5 days sampling and measuring compost of heap of heat zone every day.Fermented substrate is with germination index (GI%) measurement result, the inoculation fermentation microbial inoculum compost in the time of 14 days near 70%, and do not connect bacterium will compost in the time of 25 days just near 70%, 5 days heap temperature of the compost of inoculation fermentation microbial inoculum has reached 60 ℃, the fermented substrate comparison is 32.3% according to fast 10.2%, 25 day secondary fermentation matrix water ratio of rate of water loss, and this fermentation materials adds 10% weathered coal mixing again, as soilless culture, tomato grows fine as vegetable growing matrix.

Claims (1)

1, grey gamboge streptomycete (Streptomyces griseoluteus) W1 bacterial strain, its bacterial strain preserving number is CGMCCNO.1086.
CNB2005100979022A 2004-03-18 2004-03-18 Agricultural waste fermenting bacterial prepn and its prepn process Expired - Fee Related CN100338211C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB2005100979022A CN100338211C (en) 2004-03-18 2004-03-18 Agricultural waste fermenting bacterial prepn and its prepn process

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB2005100979022A CN100338211C (en) 2004-03-18 2004-03-18 Agricultural waste fermenting bacterial prepn and its prepn process

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN 200410039818 Division CN1264969C (en) 2004-03-18 2004-03-18 Zymogen agent for treating agricultural wastes and preparation method

Publications (2)

Publication Number Publication Date
CN1772880A true CN1772880A (en) 2006-05-17
CN100338211C CN100338211C (en) 2007-09-19

Family

ID=36760020

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB2005100979022A Expired - Fee Related CN100338211C (en) 2004-03-18 2004-03-18 Agricultural waste fermenting bacterial prepn and its prepn process

Country Status (1)

Country Link
CN (1) CN100338211C (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102010824A (en) * 2010-09-07 2011-04-13 西北农林科技大学 Microbial agent for promoting quick decomposition of agricultural wastes and preparation method
TWI422557B (en) * 2008-02-19 2014-01-11
CN108300668A (en) * 2018-01-26 2018-07-20 重庆鹓雏农业科技有限公司 Organic matter speciality fermenting agent
CN111100819A (en) * 2020-01-15 2020-05-05 北京中源创能工程技术有限公司 Bacillus and microbial agent and application thereof

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI422557B (en) * 2008-02-19 2014-01-11
CN102010824A (en) * 2010-09-07 2011-04-13 西北农林科技大学 Microbial agent for promoting quick decomposition of agricultural wastes and preparation method
CN108300668A (en) * 2018-01-26 2018-07-20 重庆鹓雏农业科技有限公司 Organic matter speciality fermenting agent
CN108300668B (en) * 2018-01-26 2021-08-17 重庆鹓雏农业科技有限公司 Organic matter special fermentation bacterial agent
CN111100819A (en) * 2020-01-15 2020-05-05 北京中源创能工程技术有限公司 Bacillus and microbial agent and application thereof

Also Published As

Publication number Publication date
CN100338211C (en) 2007-09-19

Similar Documents

Publication Publication Date Title
CN101759494B (en) Diseases-prevention organic fertilizer prepared by edible fungi residue
CN102992827B (en) Active organic bacterial fertilizer and preparation method thereof
CN103570446B (en) Process technology for producing functional fertilizer by utilizing tobacco waste
CN102286376B (en) Microbial inoculum for high-efficiency fermenting bed and preparation method thereof
CN102442849A (en) Disease-preventing bio-organic fertilizer prepared from straw and its preparation method
CN103484396B (en) New strain of streptomyces thermocarboxydus and application thereof
CN103980019A (en) Agricultural solid organic waste medium temperature biological fermentation process and application thereof
CN102337237B (en) Functional organic material decomposing agent and preparation method thereof
CN103484410B (en) Biological agent for compost treatment of livestock manure
CN103011961A (en) Active organic bacterial manure and preparation method thereof
CN101372425A (en) Microbial fertilizer and preparation thereof
CN103011916A (en) Method for preparing organic bacteria liquid, organic bacteria liquid prepared by method and application of organic bacteria liquid
CN107646616A (en) A kind of preparation method of floriculture substrate
CN104206716A (en) Biological organic aquaculture nutrition fertilizer powder and preparation method thereof
CN106305487A (en) Biological active padding material of fermentation bed for poultry, preparation method and application
CN101759493A (en) Biological disease-preventing organic fertilizer and preparation method thereof
CN1186441C (en) Microbial fertilizer bacterial agent for direct decomposing straw as fertilizer and its production process and application method
CN103173387B (en) Growth-promoting bacteria for facilitating growth of rape and microbial organic fertilizer
CN1080463A (en) Wheat straw, straw raw material bag cultivating flat mushroom high yield technique
CN106747839A (en) A kind of culture medium of edible fungus, its preparation method and application
CN1264969C (en) Zymogen agent for treating agricultural wastes and preparation method
CN108504598A (en) A kind of bio-bacterial manure leavening improving rhizosphere of strawberry soil ecology structure
CN1827762A (en) Mountain Tai aweto and artificial culture method thereof
CN100338211C (en) Agricultural waste fermenting bacterial prepn and its prepn process
CN1201000C (en) Composite cellulose decomposition bacteria and preparation and use thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20070919

Termination date: 20110318