CN105647831A - Method for accelerating growth of flocculating bacteria by using Philodina culture liquid - Google Patents
Method for accelerating growth of flocculating bacteria by using Philodina culture liquid Download PDFInfo
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- CN105647831A CN105647831A CN201610119990.XA CN201610119990A CN105647831A CN 105647831 A CN105647831 A CN 105647831A CN 201610119990 A CN201610119990 A CN 201610119990A CN 105647831 A CN105647831 A CN 105647831A
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- 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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Abstract
The invention relates to a method for accelerating growth of flocculating bacteria by using Philodina culture liquid. The Philodina culture liquid is obtained through the following steps: 1), culturing Philodina with a culture liquid that uses flour as bait; 2), when Philodina reaches 100-500 ind/mL in density, and filtering the Philodina culture liquid with a 300-mesh nylon mesh to remove Philodina from the Philodina culture liquid; 3), filtering and separating filtrate obtained in step 2) by using an ultrafiltration membrane to further remove fine particles such as flour in the filtrate so as to obtain filtrate that is the Philodina culture liquid described herein. The Philodina culture liquid described herein is used as a flocculating bacterial growth accelerator by an application amount of 0.05-1%. The method using the Philodina culture liquid to accelerate the growth of the flocculating bacteria has the advantages of ecological performance, convenience of use and significant effect.
Description
Technical field
The present invention relates to a kind of method that spinning roller worm culture fluid promotes flocculability bacterial growth.
Background technology
Activated sludge process is most widely used method in dirty water and wastewater treatment. The flocculability of activated sludge is one of key factor affecting the operation stability of active sludge processing system, treatment effeciency and effluent quality. The flocculability of activated sludge is more good, and the performance of active sludge processing system is more good. The flocculability of activated sludge depends on quantity and the activity of flocculability antibacterial, and the quantity increasing flocculability antibacterial or the activity improving flocculability antibacterial are all conducive to the raising of activated sludge flocculability.
The method improving activated sludge flocculability is segmented into two big classes: chemical method and biological methods. Chemical method is by adding inorganic flocculating agent or organic flocculant, increases the volume of activated sludge or increases the density of activated sludge, making the flocculability of activated sludge be improved. The side effect of this method is to make salinity in water increase, be easily formed secondary pollution, cause degradation problem under flocculability bacterial activity. Biological methods is by promoting that the flocculation activity of the growth of flocculability antibacterial or raising flocculability antibacterial improves the flocculability of activated sludge. Chinese invention patent (CN102816797) discloses the method by using Cys to strengthen light zymogenous bacteria excrement rhodopseudomonas flocculating property, but the method also exists the problems such as use specific and Cys the price of object is higher.
Spinning roller worm is a kind of micro-metazoa common in activated sludge, belongs to Bdelloid rotifers section on taxonomy. In activated sludge process when the Bdelloid rotifers quantity such as spinning roller worm are more, there will be the quantity of the flocculability antibacterial phenomenon that substantially flocculability many, activated sludge is substantially good in activated sludge, the active substance that its reason can be secreted with spinning roller worm containing promoting flocculability bacterial growth is relevant. Therefore, it can promote the growth of flocculability antibacterial by adding the spinning roller worm culture fluid of the active substance containing the secretion of spinning roller worm in activated sludge or flocculability bacteria culture media.
Summary of the invention
It is an object of the invention to provide a kind of method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth, the method utilizes spinning roller worm culture fluid to improve the growth of flocculability antibacterial, simple to operate, ecological, easy to use, the density of flocculability antibacterial can be increased, improve the flocculability of antibacterial, be conducive to extensive use, there are good economic benefits.
A kind of method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth, it is characterised in that concretely comprising the following steps of the method:
A. spinning roller worm is placed in spinning roller worm culture fluid and breeds, when the density of spinning roller worm reaches 100 ~ 500ind/mL, remove spinning roller worm with 300 order nylon net filters;
B. by culture fluid ultrafiltration membrance filter obtained for step a, the fine particle thing in filtrate is removed further;
C. the flocculability antibacterial that inclined-plane preserves is seeded in 100mLLB culture medium with inoculating loop, is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 24 hours, centrifugal treating culture fluid, with deionized water wash, with sterilized water, bacterial concentration is regulated to OD600Value is 0.6 ~ 0.8, and this solution is Bacteria suspension;
D. the Bacteria suspension obtained by step c is added in sterilized simulated domestic wastewater with the addition of the 5% ~ 10% of simulated domestic wastewater volume, namely obtains bacteria culture media;
E. step b gained spinning roller worm culture fluid is added in the bacteria culture media of step d gained with the addition of the 0.05% ~ 1% of bacteria culture media volume, it is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule.
Above-mentioned flocculability antibacterial is: vesicle shortwave Zymomonas mobilis, bacillus cereus, bacillus thuringiensis, bacillus subtilis, Acinetobacter bauamnnii one or more.
Above-mentioned simulated domestic wastewater formula is: 0.17g glucose, 0.16g starch, 0.233g sodium acetate, 0.025g ammonium chloride, 0.158g peptone, 0.04g Carnis Bovis seu Bubali cream, 0.0284g ammonium sulfate, 0.07g potassium dihydrogen phosphate and 0.06g sodium carbonate, and 1000mL deionized water. .
The present invention has the remarkable advantages that: 1) present invention utilizes the spinning roller worm culture fluid method to promote the growth of flocculability antibacterial, has green, environmental protection, non-secondary pollution, feature easy to use. 2) under the effect of spinning roller worm culture fluid, the growth of flocculability antibacterial obtains promotion, can make the more biological flocculant of flocculability bacterial secretory. 3) spinning roller worm culture fluid can promote the flocculability bacteria growing of multiple types, applied widely, can be effectively applied in activated sludge process.
Accompanying drawing explanation
Fig. 1 adds spinning roller worm culture fluid and does not add the dry cell weight size comparison diagram of bacillus cereus during spinning roller worm culture fluid.
Fig. 2 adds spinning roller worm culture fluid and does not add the dry cell weight size comparison diagram of vesicle shortwave Zymomonas mobilis during spinning roller worm culture fluid.
Fig. 3 adds spinning roller worm culture fluid and does not add the dry cell weight size comparison diagram of bacillus thuringiensis during spinning roller worm culture fluid.
Fig. 4 adds spinning roller worm culture fluid and does not add the dry cell weight size comparison diagram of mixed cell during spinning roller worm culture fluid.
Detailed description of the invention
Below in conjunction with accompanying drawing, embodiments of the invention are elaborated.
Embodiment 1
Utilize spinning roller worm culture fluid to promote the method that bacillus cereus (Bacilluscereus) grows, sequentially include the following steps:
(1) collecting cells of bacillus cereus: the bacillus cereus being stored in medium slant inoculating loop is seeded in LB culture medium, it is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 24 hours, centrifugal treating culture fluid, with deionized water wash, thus repeatedly after centrifugal, deionized water wash three times, with sterilized water, bacterial concentration is regulated to OD600Value 0.6. This solution is the bacteria suspension of bacillus cereus.
(2) simulated domestic wastewater preparation: weigh 0.17g glucose, 0.16g starch, 0.233g sodium acetate, 0.025g ammonium chloride, 0.158g peptone, 0.04g Carnis Bovis seu Bubali cream, 0.0284g ammonium sulfate, 0.07g potassium dihydrogen phosphate and 0.06g sodium carbonate, dissolve in 1000mL deionized water.
(3) prepared by spinning roller worm culture fluid: with the spinning roller worm in 300 order nylon net filter separation spinning roller worm culture fluid, repeatedly rinse the spinning roller worm being trapped on nylon wire 5 times with aseptic water. Spinning roller worm is transferred in sterile tap water from nylon wire, spinning roller worm density is made to reach 450ind/mL, it is subsequently placed on the shaking table of 120rpm, 24h is cultivated at 30 DEG C, use nylon net filter culture fluid, and remove the fine particle things such as antibacterial with 0.22 ��m of membrane filtration, collect filtrate, filtrate is settled to 100mL. This filtrate is spinning roller worm culture fluid.
(4) experimental group prepares: arranging two experimental grouies, one of which is matched group, and another group is spinning roller worm culture fluid interpolation group. Two experimental grouies all use 250mL conical beaker to be experiment container, and the antibacterial obtained with step (1) is for experimental subject, and the simulated domestic wastewater obtained with step (2) is for inoculum. The conical beaker of two experimental grouies adds the simulated domestic wastewater of 90mL, the bacteria suspension of 10mL respectively. The conical beaker of interpolation group adds the spinning roller worm culture fluid of 0.5mL again.
(5) antibacterial culturing: be placed on the shaking table of 120rpm by the conical beaker of two experimental grouies obtained by step (4), in 30 DEG C of continuous cultivations 48 hours.
(6) dry cell weight: adopt centrifuging to collect step (5) antibacterial cultivated, then pass through dry, weighing obtains the bacterial cell dry weight (Fig. 1) of two experimental grouies.
(7) spinning roller worm culture fluid promotes the growth result of bacillus cereus: as shown in Figure 1, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, bacillus cereus dry cell weight adds 60%, it was shown that the growth of bacillus cereus is had obvious facilitation effect by spinning roller worm culture fluid.
Embodiment 2
Utilize spinning roller worm culture fluid to promote the method that vesicle shortwave Zymomonas mobilis (Brevundimonasvesicularis) grows, sequentially include the following steps:
1. the collecting cells of vesicle shortwave Zymomonas mobilis: except strain used is vesicle shortwave Zymomonas mobilis, the preparation method of bacteria suspension is identical with the step (1) in embodiment 1.
2. simulated domestic wastewater preparation: step is identical with the step (2) in embodiment 1.
3. prepared by spinning roller worm culture fluid:
(1) spinning roller worm is inoculated into flour be bait distilled water or deionized water in.
(2) it is 120rpm in shading, shaking speed, when 30 DEG C, cultivates spinning roller worm.
(3) when the density after spinning roller worm breeds reaches 100 ~ 500ind/mL, with 300 order nylon net filter spinning roller worm culture fluid, from spinning roller worm culture fluid, spinning roller worm is removed.
(4) filtrate obtained by step (3) with ultrafiltration membrance filter again, removes further the fine particle things such as flour in filtrate, and the filtrate thus obtained is spinning roller worm culture fluid of the present invention. .
4. promote bacterial growth Preparatory work of experiment: except the bacteria suspension that bacteria used thereby suspension is vesicle shortwave Zymomonas mobilis, step is all identical with the step (4) in embodiment 1.
5. antibacterial culturing: step is identical with the step (5) in embodiment 1.
6. dry cell weight: step is identical with the step (6) in embodiment 1.
7. spinning roller worm culture fluid promotes the growth result of vesicle shortwave Zymomonas mobilis: as shown in Figure 2, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, vesicle shortwave Pseudomonad cell dry weight adds 29.7%, it was shown that the growth of vesicle shortwave Zymomonas mobilis is had good facilitation effect by spinning roller worm culture fluid.
Embodiment 3
Utilize spinning roller worm culture fluid to promote the method that bacillus thuringiensis (Bacillusthuringiensis) grows, sequentially include the following steps:
1. the collecting cells of bacillus thuringiensis: except strain used is bacillus thuringiensis, the preparation method of bacteria suspension is identical with the step (1) in embodiment 1.
2. simulated domestic wastewater preparation: step is identical with the step (2) in embodiment 1.
3. prepared by spinning roller worm culture fluid:
(1) spinning roller worm is inoculated into flour be bait distilled water or deionized water in.
(2) it is 120rpm in shading, shaking speed, when 30 DEG C, cultivates spinning roller worm.
(3) when the density after spinning roller worm breeds reaches 100 ~ 500ind/mL, with 300 order nylon net filter spinning roller worm culture fluid, from spinning roller worm culture fluid, spinning roller worm is removed.
(4) filtrate obtained by step (3) with ultrafiltration membrance filter again, removes further the fine particle things such as flour in filtrate, and the filtrate thus obtained is spinning roller worm culture fluid of the present invention. .
4. promote bacterial growth Preparatory work of experiment: except bacteria used thereby suspension is the bacteria suspension of bacillus thuringiensis, step is all identical with the step (4) in embodiment 1.
7. antibacterial culturing: step is identical with the step (5) in embodiment 1.
8. dry cell weight: step is identical with the step (6) in embodiment 1.
7. spinning roller worm culture fluid promotes the growth result of bacillus thuringiensis: as shown in Figure 2, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, vesicle shortwave Pseudomonad cell dry weight adds 67.9%, it was shown that the growth of bacillus thuringiensis is had good facilitation effect by spinning roller worm culture fluid.
Embodiment 3
The method utilizing the mixed vaccine that spinning roller worm culture fluid promotes to be mixed by bacillus cereus (Bacilluscereus), vesicle shortwave Zymomonas mobilis (Brevundimonasvesicularis), bacillus thuringiensis (Bacillusthuringiensis) these three antibacterial to grow, sequentially includes the following steps:
(1) collecting cells of mixed vaccine: bacillus cereus, vesicle shortwave Zymomonas mobilis, bacillus thuringiensis these three antibacterial are prepared into according to the step (1) in embodiment 1 respectively the bacteria suspension of single bacterium. The bacteria suspension of three kinds of bacterium is mixed into the ratio of 1:1:1 the bacteria suspension of mixed vaccine.
(2) simulated domestic wastewater preparation: step is identical with the step (2) in embodiment 1.
(3) prepared by spinning roller worm culture fluid: step is identical with the step (3) in embodiment 1.
(4) bacterial growth Preparatory work of experiment is promoted: except bacteria used thereby suspension is the bacteria suspension of mixed vaccine, step is all identical with the step (4) in embodiment 1.
(5) antibacterial culturing: step is identical with the step (5) in embodiment 1.
(6) dry cell weight: step is identical with the step (6) in embodiment 1.
Spinning roller worm culture fluid promotes the growth result of mixed vaccine: as shown in Figure 3, inoculum with the addition of the interpolation group of spinning roller worm culture fluid, compared with the matched group being not added with spinning roller worm culture fluid, mixed vaccine dry cell weight adds 52.9%, it was shown that the growth of mixed vaccine is had obvious facilitation effect by spinning roller worm culture fluid.
Claims (3)
1. one kind utilizes the method that spinning roller worm culture fluid promotes flocculability bacterial growth, it is characterised in that concretely comprising the following steps of the method:
A. spinning roller worm is placed in spinning roller worm culture fluid and breeds, when the density of spinning roller worm reaches 100 ~ 500ind/mL, remove spinning roller worm with 300 order nylon net filters;
B. by culture fluid ultrafiltration membrance filter obtained for step a, the fine particle thing in filtrate is removed further;
C. the flocculability antibacterial that inclined-plane preserves is seeded in 100mLLB culture medium with inoculating loop, is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 24 hours, centrifugal treating culture fluid, with deionized water wash, with sterilized water, bacterial concentration is regulated to OD600Value is 0.6 ~ 0.8, and this solution is Bacteria suspension;
D. the Bacteria suspension obtained by step c is added in sterilized simulated domestic wastewater with the addition of the 5% ~ 10% of simulated domestic wastewater volume, namely obtains bacteria culture media;
E. step b gained spinning roller worm culture fluid is added in the bacteria culture media of step d gained with the addition of the 0.05% ~ 1% of bacteria culture media volume, it is subsequently placed on the shaking table of 120rpm, in 30 DEG C of continuous cultivations 48 hours, namely obtain the culture fluid containing bacteria floccule.
2. according to claim 1 utilize spinning roller worm culture fluid promote flocculability bacterial growth method, it is characterised in that described flocculability antibacterial is: vesicle shortwave Zymomonas mobilis, bacillus cereus, bacillus thuringiensis, bacillus subtilis, Acinetobacter bauamnnii one or more.
3. the method utilizing spinning roller worm culture fluid to promote flocculability bacterial growth according to claim 1, it is characterized in that described simulated domestic wastewater formula is: 0.17g glucose, 0.16g starch, 0.233g sodium acetate, 0.025g ammonium chloride, 0.158g peptone, 0.04g Carnis Bovis seu Bubali cream, 0.0284g ammonium sulfate, 0.07g potassium dihydrogen phosphate and 0.06g sodium carbonate, and 1000mL deionized water.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105779345A (en) * | 2016-03-29 | 2016-07-20 | 上海大学 | Method for promoting growth of zymomonas mobilis by using philodina culture solution |
CN110894482A (en) * | 2019-12-17 | 2020-03-20 | 上海大学 | Method for promoting growth of flocculating bacteria |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
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JPH07155791A (en) * | 1993-12-09 | 1995-06-20 | Agency Of Ind Science & Technol | Method for purifying organic polluted waste water |
JPH08267094A (en) * | 1995-03-28 | 1996-10-15 | Lion Corp | Waste water treatment in production of alpha-sulfofatty acid alkyl ester |
CN101455192A (en) * | 2009-01-04 | 2009-06-17 | 上海大学 | Bdelloid rotifers mass culture method |
CN101704612A (en) * | 2009-11-19 | 2010-05-12 | 长安大学 | Integrated treatment process of enhanced flocculation and bio-contact oxidation for high-salt oil-containing wastewater |
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2016
- 2016-03-03 CN CN201610119990.XA patent/CN105647831B/en not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07155791A (en) * | 1993-12-09 | 1995-06-20 | Agency Of Ind Science & Technol | Method for purifying organic polluted waste water |
JPH08267094A (en) * | 1995-03-28 | 1996-10-15 | Lion Corp | Waste water treatment in production of alpha-sulfofatty acid alkyl ester |
CN101455192A (en) * | 2009-01-04 | 2009-06-17 | 上海大学 | Bdelloid rotifers mass culture method |
CN101704612A (en) * | 2009-11-19 | 2010-05-12 | 长安大学 | Integrated treatment process of enhanced flocculation and bio-contact oxidation for high-salt oil-containing wastewater |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105779345A (en) * | 2016-03-29 | 2016-07-20 | 上海大学 | Method for promoting growth of zymomonas mobilis by using philodina culture solution |
CN105779345B (en) * | 2016-03-29 | 2019-06-25 | 上海大学 | Promote the method for flocculability bacterial growth using spinning roller worm culture solution |
CN110894482A (en) * | 2019-12-17 | 2020-03-20 | 上海大学 | Method for promoting growth of flocculating bacteria |
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