CN110117555A - One kind being applied to naphthalene degradation bacteria and its microbial inoculum and application in sewage treatment - Google Patents
One kind being applied to naphthalene degradation bacteria and its microbial inoculum and application in sewage treatment Download PDFInfo
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- CN110117555A CN110117555A CN201910204577.7A CN201910204577A CN110117555A CN 110117555 A CN110117555 A CN 110117555A CN 201910204577 A CN201910204577 A CN 201910204577A CN 110117555 A CN110117555 A CN 110117555A
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- naphthalene
- rhodococcus
- degradation bacteria
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- corynebacteria
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
Abstract
The present invention relates to the biodegradation technique fields in sewage disposal technology, it is specially a kind of to be applied to naphthalene degradation bacteria in sewage treatment, naphthalene degradation bacteria includes bacillus subtilis, corynebacteria, Rhodococcus sp, device breeding method includes: s1, the screening of degradation bacteria, water sample is acquired from petroleum-polluted harbour, filters out bacillus subtilis, corynebacteria, Rhodococcus sp；S2, the domestication of bacterial strain.The invention also discloses one kind to be applied to naphthalene degradation bacterium preparation in sewage treatment, and the microbial inoculum includes: bacillus subtilis, corynebacteria, Rhodococcus sp, and total viable count is 1.5 × 108‑6.0×108cfu/mL.It is inoculated into filler the invention also discloses a kind of using above-mentioned naphthalene degradation bacterium preparation, filler is used in biodegradable pond be used for naphthalene application of degrading in sewage.Biological modeling of the invention has efficient quick degradation speed to naphthalene, and naphthalene degradation bacteria survival rate in high concentration sewage containing naphthalene is high, and high treating effect, processing speed is fast, and without secondary pollution.
Description
Technical field
The present invention relates to the biodegradation technique fields in sewage disposal technology, are applied at sewage more particularly to one kind
Naphthalene degradation bacteria and its microbial inoculum and application in reason.
Background technique
Naphthalene is a kind of organic compound, and the volatile crystal with peat-reek is mainly used for synthetic fuel, resin, spinning
It knits, the chemical products such as plastics, is common industrial chemicals.Naphthalene is also widely distributed in the harbour of oil pollution, soil, rubbish collection
Dissipate, etc., naphthalene is a kind of is widely present and the organic pollutant in waste water.The organic matter contained in waste water includes phenol, naphthalene, organic
Solvent etc., wherein naphthalene is common industrial chemicals, and the toxicity of naphthalene is higher, and sewage is easy smelly.Sewage containing naphthalene is commonly used at present
Processing method include: (1) physico-chemical process；(2) biochemical method；(3) chemical method.Wherein,.Physico-chemical process is handled using physical method, such as
Sedimentation layering, poor processing effect, chemical Treatment, such as using flocculant, sedimentation agent, synthetic reaction processing, cause secondary dirt
Dye, effect tears strong man's meaning open, and processing pond is easy to happen blocking, and energy consumption, equipment consumption occurs, at high cost.And biodegrade processing
Mode have low cost, high effect, advantage without secondary pollution, be current general technology means using biodegrade.
Currently, the main problem of the naphthalene in biodegrade processing sewage is: 1, the unicity problem of biological modeling, by
There is high toxicity in naphthalene, the activity of most of biological modeling is poor, needs effectively to filter out the microorganism of degradation naphthalene, together
When naphthalene certain resistance can be generated to microorganism, therefore the degradation speed and effect for reducing degradation bacteria effectively improve degradation bacteria
Time-to-live and survival activity in organic wastewater are current main problem；2, degradation bacteria is in the complex environment of sewage,
The bacterial strain of laboratory screening can decline the degrading activity of the naphthalene in sewage, can not grow or not have predetermined in complex environment
Effect；3, common degradation bacteria edman degradation Edman is to cultivate degradation bacteria in activated sludge, then passes through the new old generation of microorganism
Naphthalene is decomposed in the effect of thanking, and it is long to cultivate the time, and the screening process of degradation bacteria in the prior art is slow, screening effect is poor, and degradation bacteria
Metabolism influenced by environment such as temperature, nutrients big, treatment effect is slower, causes the processing time of sewage long.
Therefore, it is badly in need of a kind of naphthalene degradation bacteria and its microbial inoculum, the effective solution above problem.
Summary of the invention
The technical problem to be solved by the present invention is to, 1, the unicity problem of biological modeling, since naphthalene has high toxicity,
The activity of most of biological modeling is poor, needs effectively to filter out the microorganism of degradation naphthalene, while naphthalene can be to microorganism
Certain resistance is generated, the degradation speed and effect of degradation bacteria are reduced, therefore, the effective degradation bacteria that improves is in organic wastewater
Time-to-live and survival activity are current main problem；2, degradation bacteria is in the complex environment of sewage, the bacterium of laboratory screening
Strain can decline the degrading activity of the naphthalene in sewage, and scheduled effect can not be grown or do not had in complex environment；3, common
Degradation bacteria edman degradation Edman be to cultivate degradation bacteria in activated sludge, naphthalene is then decomposed by the metabolism of microorganism,
It is long to cultivate the time, and the screening process of degradation bacteria in the prior art is slow, screening effect is poor, and the metabolism of degradation bacteria
Influenced by environment such as temperature, nutrients big, treatment effect is slower, causes the processing time of sewage long.
To solve the above problems, the invention discloses:
One kind being applied to naphthalene degradation bacteria in sewage treatment, and the naphthalene degradation bacteria includes bacillus subtilis, corynebacteria, red ball
The breeding method of bacterium, the naphthalene degradation bacteria comprises the following steps that: s1, the screening of degradation bacteria, from petroleum-polluted harbour
Place's acquisition water sample, by being enriched with, purifying the degradation bacteria filtered out using naphthalene as sole carbon source；Filter out respectively bacillus subtilis,
Corynebacteria, Rhodococcus sp；S2, the domestication of bacterial strain, the bacillus subtilis filtered out, corynebacteria, Rhodococcus strain exist respectively
PH is the cultivation in the phosphate buffer of 4.5-5.8, the primary domestication to bacterial strain is realized, then by the bacterial strain filtered out in height
It is cultivated under the conditions of concentration naphthalene, realizes the secondary domestication to bacterial strain, the degradation bacteria after being tamed, degradation bacteria includes bacillus subtilis
Bacterium, corynebacteria, Rhodococcus sp.
The technical solution that the present invention further limits is the screening process of the s1 bacterial strain are as follows: the first step, the richness of strains
Collection: adding culture medium, then sterilize in culture dish, is placed in insulating box stand-by, will to acquire at petroleum-polluted harbour water
Sample places cultivation 2-8 days naturally, after growing bacterial strain, bacterial strain is transferred in culture medium using aseptic nipper, in constant temperature incubation
Culture is inverted in case, until grow strains, then save strains in 4 DEG C and be used to be inoculated with, second step, bacterial strain it is pure
Change, naphthalene degradation bacteria is purified in culture medium, the strains that the first step is obtained are inoculated into culture medium containing naphthalene, in constant incubator
Interior inversion culture is divided into 3 groups according to the content for adding naphthalene in culture medium, filters out high-concentration naphthalene and adjusts lower well-grown and can have
Effect decomposes the bacterial strain of naphthalene, and the bacterial strain filtered out continues secondarily purified；Third step, the bacterial strain of second step after purification are protected in 4 DEG C
It deposits and is used to be inoculated with.
The technical solution that the present invention further limits is the culture medium of the s1: peptone 10.0g, agar 20g, phosphoric acid
Hydrogen dipotassium 1.0g, sodium chloride 2g, glucose 1.0g add distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min.
One kind be applied to sewage treatment in naphthalene degradation bacterium preparation, the microbial inoculum include: bacillus subtilis, corynebacteria,
Rhodococcus sp, total viable count are 1.5 × 108-6.0×108cfu/mL.The bacillus subtilis, corynebacteria, Rhodococcus sp are adopted
Gained is cultivated with above-mentioned breeding method.
The technical solution that the present invention further limits is, in the microbial inoculum viable count of bacillus subtilis be 0.5 ×
108-2.5×108Cfu/mL, the corynebacteria viable count are 1 × 108-1.5×108Cfu/mL, the viable bacteria of the Rhodococcus sp
Number is 0.5 × 108-2.0×108cfu/mL。
The technical solution that the present invention further limits is, the bacterial preparation process, will be upper the following steps are included: the first step
Bacillus subtilis after purification, corynebacteria, Rhodococcus strain is stated to be seeded on test tube slant culture medium, and tried respectively
Pipe slant medium is inverted in insulating box, culture to bacterium colony is grown on culture medium, obtain bacillus subtilis, corynebacteria,
Rhodococcus strain, second step are distinguished the bacterial strain of the first step on culture transferring to fluid nutrient medium, fluid nutrient medium: peptone
10.0g, agar 20g, dipotassium hydrogen phosphate 1.0g, sodium chloride 2g, glucose 1.0g add distilled water to 1L, and 121 DEG C of high pressure sterilization,
Sterilize 20min；And its fluid nutrient medium is placed in insulating box, bacterium colony is grown in culture to culture medium, obtains bacillus subtilis
Bacterium, corynebacteria, Rhodococcus sp primary seed solution are placed in 4 degrees Celsius of preservations；Third step, by bacillus subtilis, rod-like stem
Bacterium, Rhodococcus sp primary seed solution are inoculated in fermentation medium respectively, the fermentation medium: naphthalene 1.0g, sodium chloride 0.5g, grape
Sugared 1.0g, dipotassium hydrogen phosphate 0.5g, yeast extract 0.1g add distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min；It carries out high
Density fermentation culture obtains bacillus subtilis microbial agent, corynebacteria microbial inoculum, Rhodococcus sp microbial inoculum, the 4th step, by bacillus subtilis
Bacteria agent, corynebacteria microbial inoculum, Rhodococcus sp microbial inoculum are mixed to get naphthalene degradation bacterium preparation.
The application of naphthalene degradation bacteria in a kind of sewage treatment, the naphthalene degradation bacteria are degraded naphthalene for saprobia, and use is above-mentioned
Naphthalene degradation bacterium preparation be inoculated into filler, filler is used to be used for naphthalene of degrading in sewage in biodegradable pond.
The technical solution that the present invention further limits is, is 25-35 degrees Celsius, pH 5.0-5.5 in temperature, naphthalene concentration is
100mg/L is best degradation condition, and microbial inoculum is up to 97%-98% to the degradation rate of naphthalene, this degrades to the naphthalene in sewage and waste water
Show high application value.
The technical solution that the present invention further limits is that the filler is biodegradable fiberfill, the fiber
The preparation process of filler are as follows: handle the discarded animal hair activity of recycling to obtain active keratin fiber, then by recycling
Stalk obtains active plant through activity processing, by active keratin fiber and active plant according to the ratio of 1:9-1:1
Example mixing, is added stabilizer, modifying agent, antioxidant, adhesive during mixing；It twists pressure and obtains activated fibre silk, will live
Property fiber filament is divided to two strands, Viability cordage is twined, using activated fibre rope as filler.
Stabilizer selects light stabilizer 770, light stabilizer 622, light stabilizer 944, light stabilizer UV-292 etc. wherein
It is a kind of；Certainly using other light stabilizers in the prior art can also, avoid fiber from being influenced by light source, the toughness of reinforcing fiber.
Modifying agent selects ABS, methyl methacrylate-butadiene-styrene (MBS), the one of which of polyvinyl alcohol, certainly using existing
Have other anti-impact modifiers in technology can also, avoid fiber by water impact, the impact resistance and intensity of reinforcing fiber.It is anti-
The one of which or chemical industry antioxidant such as antioxidant 245 of oxidant selection natural such as carrotene, vitamin etc.,
The one of which of antioxidant 3114, antioxidant 2246, antioxidant 300 etc., the anti-oxidant effect of the addition reinforcing fiber of antioxidant
Fruit slows down the effect that fiber is oxidized.The adhesive uses natural glue such as starch, protein, dextrin, animals and plants
One of or artificial synthesized adhesive such as polyurethane, polystyrene, polyacrylate, the ethene-vinyl acetate of glue, rosin etc.
The one of which of copolymer etc..The use of adhesive, so that keratin fiber and plant fiber degree of adhesion are high, bonding effect is good.
The technical solution that the present invention further limits is that the activity processing of the discarded animal hair of the recycling includes: elder generation
By the exhaust gas animal hair of recycling through ungrease treatment, the excess oil of hair is removed, the grease rate of the hair after ungrease treatment
2%-10%；Hair surface activity processing after ungrease treatment, surfactant is uniformly sprayed on hair, spraying process
Middle mixing, wherein surfactant is selected from polyethyleneglycol derivative, sodium n-alkylbenzenesulfonate (LAS), aliphatic alcohol polyethenoxy
The one of which of ether ammonium sulfate (AESA), sldium lauryl sulfate (K12 or SDS), lignosulfonates；It is lived after activity processing
Property keratin fiber.
The technical solution that the present invention further limits is the activity processing of the stalk of the recycling, after stalk is recycled
Even sprayed surface activating agent carries out surface-active-treatment, surfactant be polycaprolactone, dodecyl benzene sulfonic acid wherein
It is a kind of.
Compared with prior art, the invention has the benefit that
1, the unicity problem of biological modeling in the prior art is solved, naphthalene degradation bacteria of the invention is three kinds of different bacterium
Kind, difference is all had to the degradation principles of naphthalene, degradation environment etc., therefore, it is compound after naphthalene degradation bacteria degradation effect it is more preferable.2,
The domestication effect of naphthalene degradation bacteria of the invention is good, and faster to the degradation speed of naphthalene, naphthalene degradation bacteria exists the naphthalene degradation bacteria after taming
Survival activity in waste water is high, and naphthalene is not easy to generate resistance to degradation bacteria.3, composite degradation microbial inoculum of the invention keeps convenient, makes
With convenient, in effective removal waste water naphthalene, naphthalene removal rate is up to 98%.4, naphthalene degradation bacterial agent of the invention is for handling sewage
In high-concentration naphthalene, have that processing speed is fast, effect is good, without secondary pollution, noresidue, it is tasteless in pond；5, of the invention compound
The survival benefit of type naphthalene degradation agent is good, and degrading activity is not easy water environmental impact of getting dirty；6, naphthalene degradation bacterial agent of the invention is at low cost,
Use easy to spread；7, filler of the invention, it is low in cost, and the activity of naphthalene degradation agent is effectively raised, improve naphthalene drop
Survival benefit of the agent in sewage is solved, so that degradation effect is more preferable.
Specific embodiment
With reference to embodiment, the present invention is furture elucidated, it should be understood that following specific embodiments are only used for
It is bright the present invention rather than limit the scope of the invention.
Embodiment 1
One kind being applied to naphthalene degradation bacteria in sewage treatment, and naphthalene degradation bacteria includes bacillus subtilis, corynebacteria, Rhodococcus sp, naphthalene
The breeding method of degradation bacteria comprises the following steps that: s1, the screening of degradation bacteria, and water is acquired from petroleum-polluted harbour
Sample, by being enriched with, purifying the degradation bacteria filtered out using naphthalene as sole carbon source；Filter out respectively bacillus subtilis, corynebacteria,
Rhodococcus sp；S2, the domestication of bacterial strain, the bacillus subtilis filtered out, corynebacteria, Rhodococcus strain are respectively 4.5- in pH
Cultivation in 5.8 phosphate buffer realizes the primary domestication to bacterial strain, then by the bacterial strain filtered out in high-concentration naphthalene item
It is cultivated under part, realizes secondary domestication to bacterial strain, the degradation bacteria after being tamed, degradation bacteria includes bacillus subtilis, rodlike
Bacillus, Rhodococcus sp.
In the present embodiment, the screening process of s1 bacterial strain are as follows: the first step, the enrichment of strains: addition culture in culture dish
Then base sterilizes, be placed in insulating box for use, and the water sample acquired at petroleum-polluted harbour is placed cultivation 2-8 days naturally,
After growing bacterial strain, bacterial strain is transferred in culture medium using aseptic nipper, in being inverted culture in constant incubator, until growing
Strains, then strains are saved and be used to be inoculated with, second step, the purifying of bacterial strain in 4 DEG C, and naphthalene degradation is purified in culture medium
Bacterium, the strains that the first step is obtained are inoculated into culture medium containing naphthalene, in being inverted culture in constant incubator, according in culture medium
The content of addition naphthalene is divided into 3 groups, filters out high-concentration naphthalene and adjusts lower well-grown and can effectively decompose the bacterial strain of naphthalene, filters out
Bacterial strain continues secondarily purified；Third step, the bacterial strain of second step after purification save in 4 DEG C and for being inoculated with.
In the present embodiment, the culture medium of s1: peptone 10.0g, agar 20g, dipotassium hydrogen phosphate 1.0g, sodium chloride 2g,
Glucose 1.0g aggravates distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min.
Embodiment 2
One kind being applied to naphthalene degradation bacterium preparation in sewage treatment, and microbial inoculum includes: bacillus subtilis, corynebacteria, Rhodococcus sp, always
Viable count is 1.5 × 108-6.0×108cfu/mL.The bacillus subtilis, corynebacteria, Rhodococcus sp use embodiment 1
Breeding method cultivate gained.
In the present embodiment, the viable count of bacillus subtilis is 0.5 × 10 in microbial inoculum8-2.5×108Cfu/mL, it is rodlike
Bacillus viable count is 1 × 108-1.5×108Cfu/mL, the viable count of Rhodococcus sp are 0.5 × 108-2.0×108cfu/mL。
In the present embodiment, bacterial preparation process is the following steps are included: the first step, by above-mentioned bacillus subtilis after purification
Bacterium, corynebacteria, Rhodococcus strain are seeded to respectively on test tube slant culture medium, and its test tube slant culture medium is inverted in perseverance
In incubator, bacterium colony is grown in culture to culture medium, obtains bacillus subtilis, corynebacteria, Rhodococcus strain, second step will
On the bacterial strain difference culture transferring to fluid nutrient medium of the first step, fluid nutrient medium: peptone 10.0g, agar 20g, dipotassium hydrogen phosphate
1.0g, sodium chloride 2g, glucose 1.0g add distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min；And by its Liquid Culture
Base is placed in insulating box, is grown bacterium colony in culture to culture medium, is obtained bacillus subtilis, corynebacteria, Rhodococcus sp level-one kind
Sub- liquid is placed in 4 degrees Celsius of preservations；Third step connects bacillus subtilis, corynebacteria, Rhodococcus sp primary seed solution respectively
Kind is in fermentation medium, the fermentation medium: naphthalene 1.0g, sodium chloride 0.5g, glucose 1.0g, dipotassium hydrogen phosphate 0.5g, ferment
Female cream 0.1g adds distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min；High density fermentation culture is carried out, withered grass gemma is obtained
Bacillus microbial inoculum, corynebacteria microbial inoculum, Rhodococcus sp microbial inoculum, the 4th step, by bacillus subtilis microbial agent, corynebacteria microbial inoculum, red ball
Bacteria agent is mixed to get naphthalene degradation bacterium preparation.
Embodiment 3
The application of naphthalene degradation bacteria in a kind of sewage treatment, naphthalene degradation bacteria degrade naphthalene for saprobia, the naphthalene of embodiment 2 are degraded
Bacteria agent is inoculated into filler, and filler is used to be used for naphthalene of degrading in sewage in biodegradable pond.
It in the present embodiment, is 25-35 degrees Celsius, pH 5.0-5.5 in temperature, naphthalene concentration is 100mg/L, for best drop
Solution condition, microbial inoculum are up to 97%-98% to the degradation rate of naphthalene, this shows the naphthalene degradation in sewage and waste water high using valence
Value.
In the present embodiment, the filler is biodegradable fiberfill, the preparation process of the fiberfill are as follows:
It handles the discarded animal hair activity of recycling to obtain active keratin fiber, then obtains the stalk of recycling through activity processing
Active plant mixes active keratin fiber and active plant according to the ratio of 1:9-1:1, during mixing
Stabilizer, modifying agent, antioxidant, adhesive is added；It twists pressure and obtains activated fibre silk, be divided to two strands for activated fibre silk, be twined
Viability cordage, using activated fibre rope as filler.
Stabilizer selects light stabilizer 770, light stabilizer 622, light stabilizer 944, light stabilizer UV-292 etc. wherein
It is a kind of；Certainly using other light stabilizers in the prior art can also, avoid fiber from being influenced by light source, the toughness of reinforcing fiber.
Modifying agent selects ABS, methyl methacrylate-butadiene-styrene (MBS), the one of which of polyvinyl alcohol, certainly using existing
Have other anti-impact modifiers in technology can also, avoid fiber by water impact, the impact resistance and intensity of reinforcing fiber.It is anti-
The one of which or chemical industry antioxidant such as antioxidant 245 of oxidant selection natural such as carrotene, vitamin etc.,
The one of which of antioxidant 3114, antioxidant 2246, antioxidant 300 etc., the anti-oxidant effect of the addition reinforcing fiber of antioxidant
Fruit slows down the effect that fiber is oxidized.The adhesive uses natural glue such as starch, protein, dextrin, animals and plants
One of or artificial synthesized adhesive such as polyurethane, polystyrene, polyacrylate, the ethene-vinyl acetate of glue, rosin etc.
The one of which of copolymer etc..The use of adhesive, so that keratin fiber and plant fiber degree of adhesion are high, bonding effect is good.
In the present embodiment, the activity processing of the discarded animal hair of the recycling includes: the first exhaust gas animal by recycling
Hair removes the excess oil of hair, the grease rate 2%-10% of the hair after ungrease treatment through ungrease treatment；Ungrease treatment
Hair surface activity processing afterwards, surfactant is uniformly sprayed on hair, is mixed in spraying process, wherein surface
Activating agent be selected from polyethyleneglycol derivative, sodium n-alkylbenzenesulfonate (LAS), fatty alcohol polyoxyethylene ether ammonium sulfate (AESA),
The one of which of sldium lauryl sulfate (K12 or SDS), lignosulfonates；Active keratin fiber is obtained after activity processing.
The method of degreasing is the degreasing methods in the prior art such as mechanical degreasing method, enzyme process, emulsion process, solvent method, to have
The degreasing of effect, so that the grease rate of the hair after degreasing is 2%-10%.Grease rate height meeting is so that hair is sent out in subsequent use
Sour smelly (grease can decompose), grease rate is too low, and hair can be crisp easily broken.
In the present embodiment, the activity processing of the stalk of the recycling, even application surfactant after stalk is recycled
Surface-active-treatment is carried out, surfactant is polycaprolactone, the one of which of dodecyl benzene sulfonic acid.
Embodiment 4
Bacillus subtilis microbial inoculum, rod-like stem microbial inoculum, Rhodococcus sp agent and the naphthalene degradation bacterium preparation that embodiment 2 obtains are seeded to naphthalene respectively
Concentration is in the inorganic salts basic culture solution of 100mg/l, and 30 degrees Celsius, 150r/mim constant-temperature table shaken cultivation is cultivated respectively
After 1d, 3d, 5d, 7d, for carrying out degradation effect comparative test in high concentration sewage containing naphthalene.As a result such as table 1:
Table 1
Experimental data shows: when bacillus subtilis microbial inoculum, rod-like stem microbial inoculum, Rhodococcus sp agent are used alone, having good naphthalene drop
The effect of solution, bacillus subtilis microbial inoculum, the compound preparation cost invention of rod-like stem microbial inoculum, Rhodococcus sp agent naphthalene degradation bacterium preparation when,
Naphthalene degradation effect significantly improves, it is compound after effect it is more preferable, and activity is strong.Bacillus subtilis microbial inoculum, rod-like stem after cultivating 1d
Microbial inoculum, Rhodococcus sp agent cultivation and naphthalene degradation bacterium preparation show the activity of high degradation, cultivate bacillus subtilis microbial inoculum, the stick of 7d
The agent of shape bacillus, Rhodococcus sp agent cultivation and naphthalene degradation bacterium preparation, which remain unchanged, shows the activity of high degradation.
Embodiment 5
Growth conditions temperature be 25-35 degrees Celsius, pH be 5.0-5.5 under the conditions of, naphthalene concentration be 100mg/L optimum condition under,
It is inoculated with microbial inoculum amount respectively are as follows: every liter of sewage launches 0.05g microbial inoculum, 0.1g microbial inoculum, 1.5g microbial inoculum, after biodegrade for 24 hours, carries out
Degradation effect, microbial inoculum survival benefit comparative test.
Every liter of sewage launches 0.05g microbial inoculum, as a result such as table 2:
| 1h | 4h | 8h | 16h | 24h | |
| Degradation rate (%) | 62.3 | 70.1 | 82.8 | 95.9 | 98.2 |
| Survival rate (%) | 99.9 | 99.6 | 99.4 | 99.2 | 99.0 |
Table 2
Every liter of sewage launches 0.1g microbial inoculum, as a result such as table 3:
| 1h | 4h | 8h | 16h | 24h | |
| Degradation rate (%) | 62.6 | 70.3 | 83.1 | 96.2 | 98.7 |
| Survival rate (%) | 99.9 | 99.7 | 99.4 | 99.2 | 99.0 |
Table 3
Every liter of sewage launches 1.5g microbial inoculum, as a result such as table 4:
| 1h | 4h | 8h | 16h | 24h | |
| Degradation rate (%) | 65.7 | 71.2 | 83.9 | 96.6 | 98.8 |
| Survival rate (%) | 99.9 | 99.7 | 99.5 | 99.2 | 99.1 |
Table 4
The technical means disclosed in the embodiments of the present invention is not limited only to technological means disclosed in above embodiment, further includes
Technical solution consisting of any combination of the above technical features.It should be pointed out that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
Claims (10)
1. one kind is applied to naphthalene degradation bacteria in sewage treatment, it is characterised in that: the naphthalene degradation bacteria includes bacillus subtilis, stick
The breeding method of shape bacillus, Rhodococcus sp, the naphthalene degradation bacteria comprises the following steps that: s1, the screening of degradation bacteria, from by petroleum
Water sample is acquired at the harbour of pollution, by being enriched with, purifying the degradation bacteria filtered out using naphthalene as sole carbon source；Withered grass is filtered out respectively
Bacillus, corynebacteria, Rhodococcus sp；S2, the domestication of bacterial strain, the bacillus subtilis filtered out, corynebacteria, Rhodococcus sp bacterium
The strain cultivation in the phosphate buffer that pH is 4.5-5.8 respectively, realize primary domestication to bacterial strain, then will filter out
Bacterial strain is cultivated under the conditions of high-concentration naphthalene, realizes the secondary domestication to bacterial strain, the degradation bacteria after being tamed, degradation bacteria includes withered
Careless bacillus, corynebacteria, Rhodococcus sp.
2. as described in claim 1 a kind of applied to naphthalene degradation bacteria in sewage treatment, it is characterised in that: the sieve of the s1 bacterial strain
Select process are as follows: the first step, the enrichment of strains: adding culture medium, then sterilize in culture dish, is placed in insulating box for use, will
The water sample acquired at petroleum-polluted harbour is placed naturally to be cultivated 2-8 days, after growing bacterial strain, using aseptic nipper by bacterial strain
It is transferred in culture medium, in being inverted culture in constant incubator, until growing strains, then saves strains simultaneously in 4 DEG C
For being inoculated with, second step, the purifying of bacterial strain purifies naphthalene degradation bacteria in culture medium, and the strains that the first step is obtained are inoculated into
In culture medium containing naphthalene, in being inverted culture in constant incubator, filters out high-concentration naphthalene and adjust lower well-grown and can effectively decompose
The bacterial strain of naphthalene, the bacterial strain filtered out continue secondarily purified；Third step, the bacterial strain of second step after purification are used in combination in 4 DEG C of preservations
In inoculation.
3. as claimed in claim 2 a kind of applied to naphthalene degradation bacteria in sewage treatment, it is characterised in that: the culture of the s1
Base: peptone 10.0g, agar 20g, dipotassium hydrogen phosphate 1.0g, sodium chloride 2g, glucose 1.0g add distilled water to 1L, and high pressure is gone out
121 DEG C of bacterium, sterilize 20min.
4. one kind be applied to sewage treatment in naphthalene degradation bacterium preparation, it is characterised in that: the microbial inoculum include: bacillus subtilis,
Corynebacteria, Rhodococcus sp, total viable count are 1.5 × 108-6.0×108cfu/mL。
5. as claimed in claim 4 a kind of applied to naphthalene degradation bacterium preparation in sewage treatment, it is characterised in that: in the microbial inoculum
The viable count of bacillus subtilis is 0.5 × 108-2.5×108Cfu/mL, the corynebacteria viable count are 1 × 108-1.5
×108Cfu/mL, the viable count of the Rhodococcus sp are 0.5 × 108-2.0×108 cfu/ml。
6. a kind of as described in claim 4 or 5 be applied to naphthalene degradation bacterium preparation in sewage treatment, it is characterised in that: the bacterium
Agent preparation method is the following steps are included: the first step, by above-mentioned bacillus subtilis after purification, corynebacteria, Rhodococcus strain
It is seeded on test tube slant culture medium respectively, and its test tube slant culture medium is inverted in insulating box, in culture to culture medium
Bacterium colony is grown, bacillus subtilis, corynebacteria, Rhodococcus strain are obtained, the bacterial strain of the first step is distinguished culture transferring extremely by second step
On fluid nutrient medium, fluid nutrient medium: peptone 10.0g, agar 20g, dipotassium hydrogen phosphate 1.0g, sodium chloride 2g, glucose
1.0g adds distilled water to 1L, and 121 DEG C of high pressure sterilization, sterilize 20min；And its fluid nutrient medium is placed in insulating box, culture is extremely
Bacterium colony is grown on culture medium, obtains bacillus subtilis, corynebacteria, Rhodococcus sp primary seed solution, is placed in 4 degrees Celsius of guarantors
It deposits；Bacillus subtilis, corynebacteria, Rhodococcus sp primary seed solution are inoculated in fermentation medium, the hair by third step respectively
Ferment culture medium: naphthalene 1.0g, sodium chloride 0.5g, glucose 1.0g, dipotassium hydrogen phosphate 0.5g, yeast extract 0.1g add distilled water to 1L,
121 DEG C of high pressure sterilization, sterilize 20min；Carry out high density fermentation culture, obtain bacillus subtilis microbial agent, corynebacteria microbial inoculum,
Bacillus subtilis microbial agent, corynebacteria microbial inoculum, Rhodococcus sp microbial inoculum are mixed to get naphthalene degradation bacteria by Rhodococcus sp microbial inoculum, the 4th step
Microbial inoculum.
7. the application of naphthalene degradation bacteria in a kind of sewage treatment, it is characterised in that: the naphthalene degradation bacteria is used for saprobia degradation naphthalene,
It is inoculated into filler using the naphthalene degradation bacterium preparation as described in claim 4-6, filler is used to be used for dirt in biodegradable pond
It degrades in water naphthalene.
8. the application of naphthalene degradation bacteria in a kind of sewage treatment as claimed in claim 7, it is characterised in that: in temperature be 25-35
Degree Celsius, pH 5.0-5.5, naphthalene concentration is 100mg/L, is best degradation condition.
9. the application of naphthalene degradation bacteria in a kind of sewage treatment as claimed in claim 7, it is characterised in that: the filler is that can give birth to
The fiberfill of object degradation, the preparation process of the fiberfill are as follows: the discarded animal hair activity of recycling is handled and is lived
Property keratin fiber, then by the stalk of recycling through activity processing obtain active plant, by active keratin fiber and work
Property plant fiber according to 1:9-1:1 ratio mix, during mixing be added stabilizer, modifying agent, antioxidant, bonding
Agent；It twists pressure and obtains activated fibre silk, be divided to two strands for activated fibre silk, be twined Viability cordage, using activated fibre rope as filling out
Material.
10. the application of naphthalene degradation bacteria in a kind of sewage treatment as claimed in claim 7, it is characterised in that: the recycling is given up
The activity processing for abandoning animal hair includes: to remove the excess oil of hair first by the exhaust gas animal hair of recycling through ungrease treatment
It removes, the grease rate 2%-10% of the hair after ungrease treatment；Hair surface activity processing after ungrease treatment, surfactant is equal
Even is sprayed on hair, mixes in spraying process, wherein surfactant is selected from polyethyleneglycol derivative, linear alkylbenzene (LAB)
Sodium sulfonate, fatty alcohol polyoxyethylene ether ammonium sulfate, the one of which of sldium lauryl sulfate, lignosulfonates；After activity processing
Obtain active keratin fiber.
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