CN102381768A - Method for purifying mariculture wastewater by utilizing compound microbial inoculant - Google Patents
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Abstract
The invention discloses a method for purifying mariculture wastewater by utilizing a compound microbial inoculant. The method comprises the following steps: (1) screening the compound microbial inoculant which consists of photosynthetic bacteria, nitrobacteria, denitrifying bacteria, candida utilis, lactobacillus acidophilus and bacillus; (2) activating the compound microbial inoculant by utilizing a culture medium: directly adding the strain into the culture medium based on 1% of inoculation amount (w/v), and activating the strain by a shaker at the temperature of 37 DEG C with the speed of 150rpm (revolution per minute) for 24 hours so that the total content of microbes is 7*10<8>-9*10<8>CFUs (colony forming units)/mL; and (3) adding the activated compound inoculant into the treated aquaculture wastewater, wherein the compound inoculant accounts for 0.06%-0.12% of the total weight of the aquaculture wastewater. The method has the advantages of no toxicity and harm on aquaculture animals, simple process, fast organic matter degradation speed, obvious effect, no drug residues and no secondary pollution. Therefore, the method has economic feasibility and is applicable to popularization and application to coastal aquaculture industry.
Description
Technical field
The present invention relates to a kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater.
Background technology
The fast development of China's mariculture industry has driven rapid economy development in recent years, but the while has also been caused very big influence to the ecosystem of bank.Because the ecologic structure of aquaculture self and the defective of breeding way, mariculture industry has produced a series of having a strong impact on to the marine site ecosystem, as nutritive salt pollute, sedimentary environment changes, drug use is polluted, to the problems such as influence of littoral biology.Contain organism, nitrogen, phosphorus of high density etc. in the breeding wastewater, the water quality eutrophication has very big toxic action to cultured fishes.The inshore sea water culture environment pollutes and goes from bad to worse, and has had a strong impact on the aquaculture ecological security, and fishery products often suffer the body eutrophication influence and mass mortality, the Sustainable development of restriction mariculture industry.
At present, the breeding environment ecological reestablishment mainly comprises three types, and physics reparation, chemistry are repaired and biological prosthetic (Li Chunhou; Wang Xuefeng, Wang Xiaowei, Lin Lin. Chinese sea water culture environment quality and ecological reestablishment progress [J] thereof. agricultural environment science journal; 2006,25:310-315).Common physics reparation has bed mud dredging, change water, aeration, screen cloth, spill method such as zeolite ash, but these class methods often need higher investment and running expense, and treatment effect is not thorough.Chemistry reparation mainly is that utilization improver of water quality, water disinfection agent and algicide etc. improve breeding environment.But chemical renovation agent is easy to generate deleterious secondary metabolite, makes aquatic ecosystem healthy state worsen more, the fishery products quality deterioration.Biological prosthetic (Bioremediation) is meant especially microorganism catalysis degraded environmental pollutant of biology; Reduce or eliminate controlled or spontaneous process (Madsen E L.Metermining in situ biodegradation:facts and challenges [J] the .Evrion Sci Technol of environmental pollution; 1991,25 (10): 1663-1672).Utilize mikrobe apparent important, breeding environment is played the obvious effect of improving for the effect day of biological prosthetic hazardous contaminant in administering the ocean of main body.Research shows; Mikrobes such as photosynthetic bacterium, Nocardia bacteria, genus bacillus, nitrobacteria, yeast, green alga, seat algae, tenaculat Habenaria, chlorella and complex microbial community all have the good effect that purifies water; Can be used for sea farming biological prosthetic process (Liu Weidong, Su Hao, Deng Likang. the application [J] of mikrobe in aquaculture. aquatic science; 2001,20 (2): 28-31; Zhao Yujie thanks in fashionly, and week can. the application [J] of probiotics in aquaculture. and Sustainable development, 2007,1:27-29).
Chinese patent ZL 200810230123.9 discloses and has utilized the self-produced mud of sea farming system that marine culture wastewater is carried out denitrification purification, NH in the waste water
4 +-N reduces to 0.54mg/L by 0.80mg/L, NO
3 --N reduces to 41.1mg/L by 60.37mg/L.But this method is mainly removed the nitrate salt in the water body, and to organism, pollutant removals such as phosphorus are not obvious, and it is single, not thorough to handle bacterial classification.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater.The present invention is to the cultivated animals toxicological harmless, and technology is simple, and degradation speed is fast, effect is obvious, and the drug residue free problem does not cause secondary pollution.
In order to reach above-mentioned purpose, solution of the present invention is:
A kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater comprises the steps:
1) complex micro organism fungicide: form complex micro organism fungicide by photosynthetic bacterium, nitrobacteria, denitrifying bacterium, lactobacterium acidophilus, Candida utilis and genus bacillus;
2) complex micro organism fungicide adopts the substratum activation: in substratum, directly add above-mentioned complex micro organism fungicide by 1% inoculum size,, activated spawn 24h under 37 ℃ of shaking table 150rpm, making microbial total content is 7 * 10
8~9 * 10
8CFU/mL;
3) throwing of the composite fungus agent after activation bacterium amount directly adds in processing breeding wastewater 0.6 ‰~1.2 ‰ (v/v) ratio.Can adjust according to sea farming kind, density and breeding wastewater composition.
Behind five kinds of spawn culture in the described step 1) in the complex micro organism fungicide, regulate, be mixed with complex micro organism fungicide, make mikrobe total count>=1.0 * 10 in the microbial inoculum according to the composition of the marine culture wastewater that is processed
10CFU/g, and photosynthetic bacterium>=5.0 * 10 wherein
8CFU/g, nitrobacteria>=2.0 * 10
8CFU/g, denitrifying bacterium>=3.0 * 10
8CFU/g, lactobacterium acidophilus>=1.0 * 10
9CFU/g, Candida utilis>=5.0 * 10
8CFU/g, genus bacillus>=7.5 * 10
9CFU/g.
It is the red false unit cell in marsh of CCTCC M205147, the Arthrobacter of ACCC 10691, the alcaligenes dentrificans of ACCC 03247, the lactobacterium acidophilus of ACCC 10637, the Candida utilis of CGMCC 2.1016, the subtilis of CGMCC1.1630 that described five kinds of bacterial classifications adopt Chinese microorganism strain resources bank preserving number respectively; Carry out salt tolerant, heatproof domestication; As producing bacterium, form complex micro organism fungicide.
Described step 2) nutrient media components is in: peptone 3g/L, yeast powder 3g/L, glucose 5g/L, sodium-chlor 10g/L, potassium primary phosphate 0.5g/L, MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.2g/L, ammonium sulfate 1g/L, seawater 1L, transfer pH 7.0.
Beneficial effect of the present invention is: the water quality characteristics that the present invention is directed to the seawater aquiculture waste water; Utilize the potential of microbiological deterioration pollutent in the water body; Filter out the efficient degradation microorganism strains, purify water, utilize the comprehensive decomposition of synergy of various mikrobes and remove pollutent such as organism and nitrogen phosphorus in the water body through in marine culture wastewater, adding complex micro organism fungicide; Improve water quality with this, improve and culture quality; Two week of water body back COD after the processing
CrReduced more than 61.2% BOD
5Reduced more than 53.3%, total nitrogen (TN) reduces more than 38.1%, and total phosphorus reduces more than 27.4%; A safety, health, nontoxic, harmless breeding environment have been created.Complex micro organism fungicide of the present invention is to the cultivated animals toxicological harmless, and technology is simple, and degradation speed is fast, effect is obvious, and the drug residue free problem does not cause secondary pollution.Therefore, method of the present invention has economic feasibility, is adapted to promoting the use of of coastal breed industry.
Embodiment
Embodiment 1
It is the red false unit cell in marsh of CCTCC M 205147, the Arthrobacter of ACCC 10691, the alcaligenes dentrificans of ACCC 03247, the lactobacterium acidophilus of ACCC 10637, the Candida utilis of CGMCC 2.1016, the subtilis of CGMCC 1.1630 that present embodiment adopts Chinese microorganism strain resources bank preserving number; Carry out salt tolerant, heatproof domestication; As producing bacterium, form complex micro organism fungicide.Mikrobe total count>=1.0 * 10 in the complex micro organism fungicide wherein
10CFU/g, and photosynthetic bacterium>=5.0 * 10 wherein
8CFU/g, nitrobacteria>=2.0 * 10
8CFU/g, denitrifying bacterium>=3.0 * 10
8CFU/g, lactobacterium acidophilus>=1.0 * 10
9CFU/g, Candida utilis>=5.0 * 10
8CFU/g, genus bacillus>=7.5 * 10
9CFU/g.
Bacterial strain salt tolerant seed selection step is following.In containing the gradient substratum of sodium-chlor that massfraction is 0.3%-2%, cultivate the seed selection bacterial strain, progressively improve the salt resistance ability of bacterial strain.Bacterium bacterium salt resistance ability through obtaining after the domestication of several generations salt tolerant is stronger than parental strain, and has heredity.
The bacterial strain that experiment selects to salt tolerant carries out secular heatproof domestication.The heatproof domestication adopts heat shock to handle, and some has the cellular constituent of heat protective effect to induce microorganisms, thereby improves the heat resistance of mikrobe.Yeast culture thing after the activation is seeded in the MRS liquid nutrient medium, in 40,45,50,55,60 ℃ of water-baths, heats certain hour behind the cultivation certain hour, culture is placed 37 ℃ of cultivations with continued.Research shows that probiotics can obviously improve its heat impedance through the heat-inducible of certain hour, produces the survival rate of complex micro organism fungicide bacterial classification and the adaptive capacity to environment of complex microorganism thereby improve drying process with atomizing.
Adopt the substratum activation, nutrient media components is: peptone 3g/L, yeast powder 3g/L, glucose 5g/L, sodium-chlor 10g/L, potassium primary phosphate 0.5g/L, MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.2g/L, ammonium sulfate 1g/L, seawater 1L, transfer pH 7.0.In substratum, directly add above-mentioned complex micro organism fungicide by 1% inoculum size, activated spawn 24h under 37 ℃ of shaking table 150rpm, making microbial total content is 7 * 10
8~9 * 10
8CFU/mL.
The water body of present embodiment purifying treatment is sea farming base, Xiamen west marine site fish well water body A; Area is 5m * 5m; Directly add 1.2 ‰ composite bacteria liquids, after purifying treatment begins the 2nd, 4,6,8,10,12,14d measures the water quality in purifying treatment pond, water-quality guideline is COD, BOD
5, TN, TP, DO etc. (specifically measuring the result sees shown in the table 1).COD initial content in the water body is 43.6mg/L, and in the mikrobe repair process, the content of COD reduces significantly, and the content of COD is 13.8mg/L behind the original position reparation 14d, and clearance is 68.3%; BOD in the water body
5Initial content is 22.9mg/L, in the mikrobe repair process, and BOD
5Content reduce gradually, add composite bacteria 14d after, aquaculture water BOD
5Be 8.5mg/L, clearance is 62.9%; The content that the purifying treatment phase begins water body TN is 2.09mg/L, and the TN content that directly adds the composite bacteria processing reduces comparatively fast, and TN concentration maintains in the scope of a fluctuation within a narrow range then, and the highest clearance is 45.9%; TP adopts the ammonium molybdate spectrophotometry; TP initial content in the sea farming system water body is 120.1 μ g/L, directly adds in the repair process of composite bacteria processing, and TP content has reduction slightly in the water body; TP content is 88.21 μ g/L behind the 14d, and clearance is 26.6%; In the decontamination process, the DO of depth of water 0.5m is carried out in situ measurement, the initial dissolution oxygen in the sea farming system is 3.2mg/L, and maintains between 3.2~6.7mg/L.In the treating processes, the DO value changes greatly, mainly is the influence that receives temperature, and next factor such as blow, rain also has certain influence to DO content in the water body.Purifying treatment finishes, and the growth of fish well Mesichthyes is normal, has no adverse reaction.Can get thus: the complex micro organism fungicide of present embodiment is to the cultivated animals toxicological harmless, and technology is simple, and degradation speed is fast, effect is obvious, and the drug residue free problem does not cause secondary pollution.
Embodiment 2
It is the red false unit cell in marsh of CCTCC M 205147, the Arthrobacter of ACCC 10691, the alcaligenes dentrificans of ACCC 03247, the lactobacterium acidophilus of ACCC 10637, the Candida utilis of CGMCC 2.1016, the subtilis of CGMCC 1.1630 that present embodiment adopts Chinese microorganism strain resources bank preserving number; Carry out salt tolerant, heatproof domestication (condition is with embodiment 1); As producing bacterium, form complex micro organism fungicide.Mikrobe total count>=1.0 * 10 in the complex micro organism fungicide wherein
10CFU/g, and photosynthetic bacterium>=5.0 * 10 wherein
8CFU/g, nitrobacteria>=2.0 * 10
8CFU/g, denitrifying bacterium>=3.0 * 10
8CFU/g, lactobacterium acidophilus>=1.0 * 10
9CFU/g, Candida utilis>=5.0 * 10
8CFU/g, genus bacillus>=7.5 * 10
9CFU/g.
Adopt the substratum activation, nutrient media components is: peptone 3g/L, yeast powder 3g/L, glucose 5g/L, sodium-chlor 10g/L, potassium primary phosphate 0.5g/L, MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.2g/L, ammonium sulfate 1g/L, seawater 1L, transfer pH 7.0.In substratum, directly add above-mentioned complex micro organism fungicide by 1% inoculum size, activated spawn 24h under 37 ℃ of shaking table 150rpm, making microbial total content is 7 * 10
8~9 * 10
8CFU/mL.
The water body of present embodiment purifying treatment is sea farming base, Xiamen west marine site fish well water body B; Area is 5m * 5m; Directly add 0.8 ‰ composite bacteria liquids, after purifying treatment begins the 2nd, 4,6,8,10,12,14d measures the water quality in purifying treatment pond, water-quality guideline is COD, BOD
5, TN, TP, DO etc. (specifically measuring the result sees shown in the table 2).Sea farming system COD in water body initial content is 34.8mg/L, and in the mikrobe repair process, the content of COD reduces gradually, and the content of COD is 10.7mg/L behind the original position reparation 14d, and clearance is 69.2%; BOD in the water body
5Initial content is 19.6mg/L, add composite bacteria 14d after, aquaculture water BOD
5Be 7.3mg/L, clearance is 62.8%; The content that the processing phase begins water body TN is 2.11mg/L, and the TN that directly adds the composite bacteria processing significantly reduces, and the highest clearance is 46.9%; TP initial content in the sea farming system water body is 105.9 μ g/L, directly adds in the repair process of composite bacteria processing, and TP content has to a certain degree reduction in the water body, and TP content is 70.60 μ g/L after the 14d, and clearance is 33.3%; In the treating processes, the DO of depth of water 0.5m is carried out in situ measurement, the initial dissolution oxygen in the sea farming system is 3.9mg/L, and in the decontamination process, the DO value fluctuates between 3.9~6.6mg/L.Purifying treatment finishes, and the growth of purifying treatment pond Mesichthyes is normal, has no adverse reaction.Can get thus: the complex micro organism fungicide of present embodiment is to the cultivated animals toxicological harmless, and technology is simple, and degradation speed is fast, effect is obvious, and the drug residue free problem does not cause secondary pollution.
Comparative Examples 1
The water body of present embodiment is sea farming base, Xiamen west marine site fish well water body A; Area is 5m * 5m, does not add the compound thing microbial inoculum of supporting one's family, and keeps normal cultivation density; The 2nd, 4,6,8,10,12,14d measures the water quality in pond, water-quality guideline is COD, BOD
5, TN, TP, DO etc. (specifically measuring the result sees shown in the table 3).COD initial content in the sea farming system water body is 42.1mg/L, and the content of COD is 40.2mg/L behind the 14d, and the content amplitude of COD is little, only reduces by 4.5%; BOD in the water body
5Initial content is 20.7mg/L, the BOD in the water body
5To fall appear in content-liter-trend of falling, aquaculture water BOD behind the 14d
5Detected value is 18.2mg/L, reduces by 12.1%; The content that the monitoring phase begins water body TN is 2.03mg/L, and TN concentration has reduction by a small margin in the observation process, raises again then, and the content of TN is 2.06mg/L behind the 14d; TP initial content in the sea farming system water body is 115.3 μ g/L, and first the reduction afterwards of TP concentration raises in the observation process, and TP content is 129.1 μ g/L after the 14d; Initial DO in the sea farming system is 3.5mg/L, and in the mensuration process, the DO value raises earlier and afterwards reduces, and maintains between 3.5~5.7mg/L.
Comparative Examples 2
The water body of present embodiment is sea farming base, Xiamen west marine site fish well water body B; Area is 5m * 5m, does not add composite bacteria liquid, keeps normal cultivation density; The 2nd, 4,6,8,10,12,14d measures the water quality of handling the pond, water-quality guideline is COD, BOD
5, TN, TP, DO etc. (specifically measuring the result sees shown in the table 4).COD initial content in the sea farming system water body is 35.3mg/L, and the content of observation process COD is fluctuation up and down in more among a small circle, and the content of COD is 34.7mg/L behind the 14d, and clearance is 1.7%; BOD in the water body
5Initial content is 18.9mg/L, BOD behind the 14d
5Be 17.6mg/L, clearance is merely 6.9%; The content that detection period begins water body TN is 1.95mg/L, TN concentration fluctuation up and down within the specific limits in the process, and the content of TN is 1.92mg/L behind the 14d; TP initial content in the sea farming system water body is 110.1 μ g/L, and TP content is 105.2 μ g/L behind the 14d; Initial DO in the sea farming system is 4.2mg/L, and in the testing process, the DO value maintains between 3.6~5.2mg/L.
Can get by the foregoing description: adopt complex micro organism fungicide purifying sea water breeding wastewater of the present invention, can the synergy through various mikrobes decompose and remove pollutents such as organism and nitrogen phosphorus in the water body, improve water quality, improve the breed quality with this; Two week of water body back COD after the processing
CrReduced more than 61.2% BOD
5Reduced more than 53.25%, total nitrogen reduces more than 38.12%, and total phosphorus reduces more than 27.40%; A safety, health, nontoxic, harmless breeding environment have been created.
Table 1
Table 2
Table 3
Table 4
Claims (4)
1. a method of utilizing complex micro organism fungicide purifying sea water breeding wastewater is characterized in that comprising the steps:
1) screening complex micro organism fungicide: form complex micro organism fungicide by photosynthetic bacterium, nitrobacteria, denitrifying bacterium, lactobacterium acidophilus, Candida utilis and genus bacillus;
2) complex micro organism fungicide adopts the substratum activation: in substratum, directly add above-mentioned complex micro organism fungicide by 1% inoculum size (w/v), and activated spawn 24h under 37 ℃ of shaking table 150rpm, making microbial total content is 7 * 10
8~9 * 10
8CFU/mL;
3) throwing of the composite fungus agent after activation bacterium amount adds in processing breeding wastewater 0.6 ‰~1.2 ‰ ratios.
2. a kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater as claimed in claim 1; It is characterized in that: behind five kinds of spawn culture in the described step 1) in the complex micro organism fungicide; Complex micro organism fungicide is regulated, is mixed with to composition according to the marine culture wastewater that is processed, makes mikrobe total count>=1.0 * 10 in the microbial inoculum
10CFU/g, and photosynthetic bacterium>=5.0 * 10 wherein
8CFU/g, nitrobacteria>=2.0 * 10
8CFU/g, denitrifying bacterium>=3.0 * 10
8CFU/g, lactobacterium acidophilus>=1.0 * 10
9CFU/g, Candida utilis>=5.0 * 10
8CFU/g, genus bacillus>=7.5 * 10
9CFU/g.
3. a kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater as claimed in claim 2; It is characterized in that: it is the red false unit cell in marsh of CCTCC M 205147, the Arthrobacter of ACCC 10691, the alcaligenes dentrificans of ACCC 03247, the lactobacterium acidophilus of ACCC 10637, the Candida utilis of CGMCC 2.1016, the subtilis of CGMCC 1.1630 that described five kinds of bacterial classifications adopt Chinese microorganism strain resources bank preserving number respectively; Carry out salt tolerant, heatproof domestication; As producing bacterium, form complex micro organism fungicide.
4. a kind of method of utilizing complex micro organism fungicide purifying sea water breeding wastewater as claimed in claim 1; It is characterized in that described step 2) in nutrient media components be: peptone 3g/L, yeast powder 3g/L, glucose 5g/L, sodium-chlor 10g/L, potassium primary phosphate 0.5g/L, MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.2g/L, ammonium sulfate 1g/L, seawater 1L, transfer pH 7.0.
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