CN101386821B - Special effect ammonifiers and waste water processing method using thereof - Google Patents
Special effect ammonifiers and waste water processing method using thereof Download PDFInfo
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- CN101386821B CN101386821B CN2007100769593A CN200710076959A CN101386821B CN 101386821 B CN101386821 B CN 101386821B CN 2007100769593 A CN2007100769593 A CN 2007100769593A CN 200710076959 A CN200710076959 A CN 200710076959A CN 101386821 B CN101386821 B CN 101386821B
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- bacterial strain
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- ammonifying bacteria
- ammonifier
- hja07
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Abstract
The invention discloses a specific ammonifier. The strain is an ammonifier strain HJA07, wherein the preserving number of the ammonifier strain is CGMCC No. 2162 and the preserving date of the ammonifier strain is September 13th, 2007. The ammonifier strain is separated from sludge, cultured in an enriched medium and separated in a separation culture medium, wherein the pH value of the ammonifierat a temperature of between 25 and 35 DEG C is between 6.5 and 8.0. By adopting the ammonifier to treat waste water, the cost is low, the efficiency is stable and no secondary pollution produces, thereby effectively treating waste water which contains organic nitrogen and protecting the environment.
Description
[technical field]
The present invention relates to biological technical field, especially about a strain special efficacy ammonifying bacteria and handle the method for waste water.
[background technology]
Nitrogen in the water body mainly exists with organonitrogen and two kinds of forms of inorganic nitrogen.The former has protein, polypeptide, amino acid and urea etc., and they derive from trade effluent (as butchering processing, printing and dyeing, process hides etc.), sanitary wastewater and agricultural wastes (straw, excrement of animals etc.) etc.Nitrate pollution not only can cause body eutrophication, causes the growth of waterplant and algae excessive, water quality is worsened rapidly, and can reduce the water body ornamental value, endangers hydrobiological existence.
[summary of the invention]
Technical problem to be solved by this invention is, overcomes the deficiencies in the prior art, provides that a kind of cost is low, the special efficacy ammonifying bacteria of removing ammonia nitrogen stabilised efficiency, non-secondary pollution and the method for handling waste water.。
The technical solution adopted for the present invention to solve the technical problems is: a strain special efficacy ammonifying bacteria, and this bacterial strain is HJA07, and ammonifying bacteria bacterial strain, preserving number are CGMCC № 2162, and preservation date is on 09 13rd, 2007.
Described ammonifying bacteria bacterial strain separates from mud and obtains, and this ammonifying bacteria is 25 ℃~35 ℃, and the pH value is 6.5~8.0, cultivates in the enrichment medium, separates in the isolation medium.
Described ammonifying bacteria bacterial strain separates from bed-silt and obtains, and it is 28 ℃ that this ammonifying bacteria bacterial strain suits in temperature, and the pH value is 7.2~7.4, cultivates in the enrichment medium, separates in the isolation medium.
Described enrichment culture based component is: peptone 10g, sal epsom 0.5g, dipotassium hydrogen phosphate 1g, sodium-chlor 0.5g, ferrous sulfate 0.01g, liquid microelement 1.0ml, distilled water 1000ml is 7.2~7.4 with 10% yellow soda ash adjust pH.
Described separation and Culture based component is: extractum carnis 3g, peptone 10g, sodium-chlor 5g, potassium primary phosphate 15mg, distilled water to 1000ml, pH value are 7.2~7.4.
Described ammonifying bacteria bacterial strain separates from bed-silt and obtains, and it is 28 ℃ that this ammonifying bacteria bacterial strain suits in temperature, and shaking culture is 2 days on the shaking table of 150r/min; In described enrichment medium, cultivate; When temperature is 28 ℃, in described isolation medium, separate again.
Described liquid microelement is: boric acid 0.5g, Sodium orthomolybdate 0.5g are dissolved in the 100ml distilled water.
Described ammonifying bacteria bacterial strain belongs to micrococcus sp (Micrococcus), is micrococcus luteus (Micrococcus luteus), and principal character is: colonial morphology is circular, yellow, smooth, low protruding, opaque, diameter 2.5mm; The bacterial strain individuality is spherical, and diameter is 0.9~1.8 μ m.
The physiological and biochemical property of described ammonifying bacteria bacterial strain shows as: Gram-positive, and rare motion, the spore of not sprouting contacts mould, oxidase positive, can grow in 5% NaCl, and the G+C mol% of DNA is 64%~75%, and organonitrogen is converted into ammonia nitrogen.
A kind ofly utilize a described strain special efficacy ammonifying bacteria to handle the method that contains organonitrogen sewage, comprise the steps:
A) cultivation of ammonifying bacteria bacterial strain;
B) separation of ammonifying bacteria bacterial strain;
C) reaction of ammonifying bacteria bacterial strain and sewage removes organonitrogen;
D) reclaim the ammonifying bacteria bacterial strain, recycle.
The invention has the beneficial effects as follows, handle waste water by ammonifying bacteria, cost is low, stabilised efficiency, non-secondary pollution, contains organic ammonia waste water thereby handled effectively, the protection environment.
[description of drawings]
Fig. 1 is the influence curve figures of different pH to the HJA07 bacterial strain;
Fig. 2 is the influence curve figure of differing temps to the HJA07 bacterial strain;
Fig. 3 is a HJA07 bacterial strain ammonification ability measurement result histogram;
Fig. 4 is a HJA07 bacterium river sewage ammonification rate diagram graphic representation.
[embodiment]
A kind of special efficacy Ammonifying bacteria of the present invention belongs to micrococcus sp (Micrococcus), is micrococcus luteus (Micrococcus luteus), called after HJA07.Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on 09 13rd, 2007, preserving number is CGMCC № 2162.
A kind of special efficacy Ammonifying bacteria of the present invention, the HJA07 bacterial strain is taken from Shenzhen's Buji bed-silt, obtains the strong bacterial strain of a strain ammonification ability through 4 enrichment culture, 3 separation and purification.Its colony characteristics is: colonial morphology is circular, yellow, smooth, low protruding, opaque, diameter 2.5mm; The bacterial strain individuality is spherical, and diameter is 0.9~1.8 μ m.The major microorganisms feature shows as: Gram-positive, and rare motion, the spore of not sprouting contacts mould, oxidase positive, can grow in 5% NaCl, and the G+C mol% of DNA is 64%~75%.Can find out that from Fig. 1, Fig. 2 HJA07 ammonifying bacteria optimum growing condition is: pH 6.5~8.0,25~35 ℃ of temperature.
A kind of special efficacy Ammonifying bacteria of the present invention, HJA07 can carry out aminating reaction under the oxygen condition of holding concurrently, can effectively organonitrogen be converted into ammonia nitrogen, will play a significant role in the dirty biological denitrificaion practice of water.
A kind of special efficacy Ammonifying bacteria of the present invention, the removal of nitrogen comprises processes such as ammonification, nitrification and denitrification in the biological denitrificaion: ammonification is to utilize Ammonifying bacteria that organonitrogen is converted into the process of ammonia-state nitrogen, and nitrification is oxidized to nitrite and nitrate by nitrite bacterium and nitrate bacterium with ammoniate; Denitrification also claims denitrogenation, under anaerobism or little anaerobic condition nitrate reduction is become Nitrous Oxide or nitrogen by denitrifying bacterium.Ammonification is the initial link in the biological denitrificaion link, is directly connected to follow-up denitrification process, and therefore, the ability to function that how to improve Ammonifying bacteria is to guarantee the biological denitrificaion important factor to strengthen ammonification.The core of ammonification is an Ammonifying bacteria, is a class functional flora important in the nature Nitrogen Cycling, in the water body ammonifying bacteria quantity and activity directly affect water quality condition.The ability of different its ammonifications of Ammonifying bacteria is also variant, and its growth conditions is also different, and the screening and separating from physical environment goes out efficiently bacterial strain, and to be applied to that enhancing sewage biological handles be the important means that improves sewage disposal usefulness.Exist multifarious microorganism species in the aquatic ecosystem such as river course, lake, circulation, migration, conversion and delay to nitrogen play an important role.Therefore, to the Ammonifying bacteria in the mud at the bottom of the river course screen, separation and purifying be expected to obtain ammonification bacterial strain efficiently, for improving bio-denitrifying sewage efficient important meaning arranged.
Embodiment 1: the mensuration of the separation of Ammonifying bacteria HJA07, purifying and physio-biochemical characteristics
(1) collected specimens
Collection in worksite Shenzhen Buji bed-silt sample is smashed with glass strain vibration behind the adding distil water in the aseptic triangular flask of 500ml.
(2) bacterial strain enrichment, separation
Extracting sample solution 10ml is to the triangular flask that 150ml enrichment medium (sterilizing) is housed, and shaking culture is 2 days on 28 ℃, the shaking table of 150r/min.Enrichment culture based component: peptone 10g, MgSO
40.5g, K
2HPO
41g, NaCl 0.5g, FeSO
40.01g, liquid microelement 1.0ml (liquid microelement: boric acid 0.5g, Sodium orthomolybdate 0.5g is dissolved in the 100ml distilled water), distilled water 1000ml, regulating the pH value with 10% yellow soda ash is 7.2~7.4.After substratum became muddiness, transferase 45 ml was to new enrichment medium (150ml) again, and shaking culture is 2~3 days under the similarity condition, repeats so 3 times again, and each transfer amount is respectively 3ml, 1ml, 0.5ml.After the bacteria suspension of getting 1ml is diluted to 10000 times, get the dull and stereotyped coating of dilution bacterium liquid 0.5ml, cultivated 2~3 days in 28 ℃, promptly have bacterium colony to occur.Select the different bacterium colony of those forms to separate, repeat to separate more than 3 times obtaining the pure bacterial strain of many strains according to the purebred isolating ordinary method plate streaking of microorganism.The separation and Culture based component is: extractum carnis 3g, peptone 10g, NaCl 5g, KH
2PO
415mg, distilled water are to 1000ml, and the pH value is 7.2~7.4.Its colony characteristics is: be faint yellow, the lawn on the solid medium is circular, and bacterium colony is less, is papillary, and edge clear is faint yellow, and glossiness is less, does not thickness, and is opaque.Diameter 2.5mm, the bacterial strain individuality is spherical, and diameter is 0.9~1.8 μ m.
(3) mensuration of Ammonifying bacteria HJA07 bacterial strain physio-biochemical characteristics
To separate, HJA07 bacterial strain behind the purifying, utilize the micro-biochemical plate of SMF (A) biochemical analysis of being correlated with.Analysis indexes comprises that mainly carbohydrate produces sour experiment, hydrolysising experiment, gramstaining experiment, anaerobic growth experiment, catalase experiment, oxydase experiment, esterase (Tween80) experiment, VP experiment, utilizes Citrate trianion experiment, 7.5%NaCl growth experiment, N.F,USP MANNITOL experiment, gelatin hydrolysate experiment, MR test and nitrate reduction experiment (method is seen " common bacteria system identification handbook "), the results are shown in Table 1.
The physiological and biochemical property of table 1.HJA07
The mensuration of embodiment 2:HJA07 bacterial strain ammonification ability
With the HJA07 bacterial strain behind the purifying, insert in the 150ml enrichment culture liquid shaking culture 48h under 28 ℃, 150r/min.The bacteria suspension of getting above-mentioned HJA07 is inoculated in the new enrichment medium of 12 200ml by 2% inoculum size, other establishes one bottle of contrast, lucifuge, vibration, 28 ℃ of cultivations, respectively took out three samples then respectively at 6,12,18,24 hours, through the centrifugal 30min of 5000r/min, get supernatant liquor with nessler reagent colorimetric method for determining ammonia nitrogen, and with the ammonia nitrogen value contrast of the nutrient solution that does not connect bacterium, calculate ammonification speed.Ammonification speed=(A
N-B
N)/t; A wherein
N: connect the ammonia nitrogen (mg/L) of the enrichment culture liquid of bacterium, B
N: the ammonia nitrogen (mg/L) after the enrichment culture liquid that does not connect bacterium is handled, incubation time (h).Measure the ammonification ability of different time.As Fig. 3, the ammonification speed of HJA07 bacterial strain 18h is 18.79mg/ (Lh), and ammonia-nitrogen content is 398.3mg/L, is converted into ammonia nitrogen substantially at 24 hours organonitrogens.Embodiment 3:HJA07 bacterial strain is handled the river sewage test
Gather Buji river, Shenzhen sewage, after measured NH
4-N content 19.7mg/L, (cell concentration is 10 to get the HJA07 bacteria suspension of 1000ml disinfection inoculation 10ml
8CFU/L), establish three parallel, other establishes one bottle and compares, in 28 ℃, 150r/min shaking culture.The variation of ammonia-nitrogen content and speed (as table 2, Fig. 4).
Table 2HJA07 bacterial strain is handled the river sewage test-results
Above content be in conjunction with concrete preferred implementation to further describing that the present invention did, can not assert that concrete enforcement of the present invention is confined to these explanations.For the general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (2)
1. a strain special efficacy ammonifying bacteria, it is characterized in that: this bacterial strain is micrococcus luteus (Micrococcus luteus) HJA07 in the micrococcus sp, and ammonifying bacteria bacterial strain, preserving number are CGMCC № 2162.
2. one kind is utilized the described special efficacy ammonifying bacteria of claim 1 to handle the method that contains organonitrogen sewage, it is characterized in that: comprise the steps:
A) cultivation of ammonifying bacteria bacterial strain;
B) separation of ammonifying bacteria bacterial strain;
C) reaction of ammonifying bacteria bacterial strain and sewage removes organonitrogen;
D) reclaim the ammonifying bacteria bacterial strain, recycle.
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陈赵芳等.一株异养硝化菌的筛选及其脱氮条件.东南大学学报(自然科学版)37 3.2007,37(3),486-490. |
陈赵芳等.一株异养硝化菌的筛选及其脱氮条件.东南大学学报(自然科学版)37 3.2007,37(3),486-490. * |
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