CN104312963A - Method for separating purified bdellovibrio from active sludge - Google Patents

Method for separating purified bdellovibrio from active sludge Download PDF

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CN104312963A
CN104312963A CN201410587775.3A CN201410587775A CN104312963A CN 104312963 A CN104312963 A CN 104312963A CN 201410587775 A CN201410587775 A CN 201410587775A CN 104312963 A CN104312963 A CN 104312963A
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bdellovibrio
separation
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active sludge
host bacteria
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余冉
李传扬
张诗文
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Southeast University
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Abstract

The invention discloses a method for separating purified bdellovibrio from active sludge. By virtue of key techniques of host bacteria screening and active sludge pretreatment and technical steps such as preparation of a culture medium and a culture solution and optimization of culture conditions and the like, efficient bdellovibrio is separated and purified from a homologous sludge sample by optimization of a series of key steps by virtue of gram negative bacteria which are obtained in active sludge as host bacteria for separating and purifying bdellovibrio. The invention not only puts forwards a method for successfully separating the efficient bdellovibrio from the active sludge of a municipal wastewater treatment plant for the first time, but also the culture period of the method is remarkably shortened, so that the defects that the existing conventional method is unstable in bdellovibrio separating effect, long in culture period, tedious in operation and the like.

Description

A kind of method of separation and purification Bdellovibrio from active sludge
 
Technical field
The invention belongs to microbial technology field, relate to the optimization method of a kind of separation and purification Bdellovibrio from municipal sewage plant second pond active sludge.
 
Background technology
Bdellovibrio class biology ( bdellovibrio-and-like organisms, be called for short Bdellovibrio) be the small-sized parasitics bacterium that a class is made a living to prey on host bacteria, Bdellovibrio individuality is less, wide 0.2 ~ 0.6 μm, long 0.5 ~ 2.0 μm, can through the millipore filtration of 0.45 μm, thalline is arcuation or shaft-like, be a kind of aerobic gram negative bacterium, it can be invaded addicted to suspended bacterial, has very high lytic activity to the gram negative bacterium of section most of in environment, genus and minority gram positive bacterium.Therefore the unique ecological advantage that has of Bdellovibrio, can be considered one of important biomolecule factor that naturally purifies, and can be applicable to the aspects such as Disease ecology control, agricultural planting, herding and aquaculture, purification of water quality, the monitoring of people and animals' hygienic safety.But from municipal sludge, be separated at present Bdellovibrio rarely have report for the research of sewage treatment plant tail water process and mud decrement .therefore, be necessary that the separation from municipal sludge first finding a kind of stability and high efficiency has the method for the Bdellovibrio of phagocytosis.
Conventional Bdellovibrio separation method can be subject to the impact of many factors in operation, as nutritive ingredient, Ca contained in the concentration of agar powder in inappropriate sample pretreatment means, substratum, substratum 2+and Mg 2+deng the concentration of trace element, pH and selected host bacteria and concentration, culture temperature etc. when being separated Bdellovibrio, if condition control improper, not only can the Extending culture cycle, even can cause the failure that Bdellovibrio is separated.After improving one's methods of relating in the present invention is through experimental exploring repeatedly, the method system of separation and purification Bdellovibrio from municipal sludge of the complete set formed gradually, comprises the pre-treatment of sample and the optimization of every culture condition and separation and purification.By adopting present method, researchist is repeatedly separated and obtains efficient Bdellovibrio from different mud sample.
 
Summary of the invention
technical problem:the present invention fully takes into account the impact of various factors in Bdellovibrio sepn process, provides a kind of method that can obtain stability and high efficiency Bdellovibrio, efficient separation and purification Bdellovibrio from active sludge.
technical scheme:the method of the present invention high efficiency separation Bdellovibrio from active sludge, comprises the following steps:
1) preparation of screening and separating Host Strains and bacteria suspension thereof in mud sample: first using the active sludge of municipal sewage plant second pond as mud sample, therefrom separate Gram-negative bacteria, then using described Gram-negative bacteria as host bacteria, after liquid propagation 18 ~ 24h is carried out to it, obtain bacterial plaque through centrifugation; Finally the resuspended host bacteria suspension that obtains is carried out to obtained bacterial plaque sterile phosphate buffer, it is preserved at 4 DEG C;
2) pre-treatment is carried out to obtain Bdellovibrio stoste to mud sample, idiographic flow is: by the homology mud sample vortex oscillation 30 ~ 60min of screening Host Strains, after room temperature leaves standstill 20 ~ 30min, utilize high speed freezing centrifuge 4 ~ 10 DEG C, rotating speed be 800 × g ~ 1000 × g condition under centrifugal 15 ~ 25min, get supernatant liquor, then by supernatant liquor 4 ~ 10 DEG C, rotating speed be 15000 × g ~ 30000 × g under centrifugal 20 ~ 30min, get precipitation, and resuspended with sterile phosphate buffer, the re-suspension liquid that obtains is Bdellovibrio stoste;
3) separation of Bdellovibrio is carried out according to following flow process, then according to this flow process, three purifying are at least repeated to the mixed solution obtained, obtain pure Bdellovibrio: first adopt the double-deck agar plate method of dilution nutrient broth to carry out the separation of Bdellovibrio, then join in dilution nutrient broth nutrient solution by being separated the Bdellovibrio obtained, and add the host bacteria suspension that described step 1) obtains wherein, its volume accounts for 0.5% ~ 1% of dilution nutrient broth nutrient solution, after vortex oscillation mixing, be transferred to 25 ~ 30 DEG C, in the constant temperature water bath shaking table of 150 ~ 200r/min, liquid increases 3 ~ 5 days, by proliferating liquid gradient dilution 10 5~ 10 7doubly, get each gradient dilution liquid and mix with host bacteria suspension,
4) preparation of pure Bdellovibrio suspension: the bacterial plaque of the pure Bdellovibrio described step 3) obtained joins in Sterile dilution nutrient broth nutrient solution, and add the host bacteria suspension that described step 1) obtains wherein, its volume accounts for 0.5% ~ 1% of dilution nutrient broth nutrient solution, be transferred to 25 ~ 30 DEG C, cultivate 3 ~ 5 days in the constant temperature water bath shaking table of 150 ~ 200r/min, the proliferating liquid obtained is pure Bdellovibrio suspension.
In the preferred version of the inventive method, the idiographic flow adopting the double-deck agar plate method of dilution nutrient broth to carry out the separation of Bdellovibrio in step 3) is: carry out gradient dilution to Bdellovibrio stoste, get each gradient dilution liquid of Bdellovibrio and host bacteria suspension mixes, and add the top-layer agar substratum of 3 ~ 7mL insulation between 50 ~ 55 DEG C wherein, after vortex oscillation mixing, evenly be laid on the prefabricated bottom agar substratum solidified, the constant temperature biochemical cultivation case putting into 25 ~ 30 DEG C after top-layer agar culture medium solidifying is cultivated 2 ~ 4 days.
In the preferred version of the inventive method, in step 3), the pH of the nutrient agar used in all separation and purification processes and dilution nutrient broth nutrient solution all needs to regulate between 6.8 ~ 7.5.
In the preferred version of the inventive method, in step 3), in the nutrient agar used in double-deck agar plate method, also comprise the CaCl that mass concentration is 0.25 ‰ ~ 0.35 ‰ 2, mass concentration be 0.40 ‰ ~ 0.50 ‰ MgCl 2.
In the preferred version of the inventive method, the concentration of the host bacteria suspension used in step 3) and step 4) is 10 8~ 10 12cfu/mL, in described step 3), the concentration ratio that Bdellovibrio gradient dilution liquid mixes with host bacteria suspension is 1/10 ~ 1/100.
The inventive method is to dilute based on the double-deck agar plate method of nutrient broth, whole sepn process relates to the separation screening of host bacteria, the pre-treatment of mud sample, the preparation of double-deck nutrient agar and the Optimal improvements of culture condition, and the isolation and purification of Bdellovibrio.The employing of the inventive method, provide not only the approach screening efficient Bdellovibrio from municipal sludge, and effectively simplifies operation steps, improves the separating effect of Bdellovibrio on double-layer plate, significantly shorten its culture cycle.
beneficial effect:compared with prior art, the present invention has the following advantages:
Content of the present invention comprises the adjustment and optimisation of correlative factor in the specific pre-treatment of screening and separating host bacteria, mud sample in municipal sewage plant active sludge, the allotment of substratum and culturing process, until the isolation and purification of Bdellovibrio.
The municipal sewage plant second pond active sludge that Bdellovibrio in the present invention is identical from source with the equal screening and separating of host bacteria, for Bdellovibrio provides the host bacteria of nutritive ingredient and the homology of Bdellovibrio to substantially reduce the time of lag phase in Bdellovibrio growth cycle, be conducive to Bdellovibrio and adapt to culture environment rapidly, acceleration its own amplification grows, thus effectively shorten culture cycle, and the separation of routine normally buys a kind of specific host bacterium from research institution, seldom have and screen from source the separation that host bacteria carries out Bdellovibrio, Bdellovibrio will be caused like this to have extremely strong dependency to a certain host bacteria, once leave this bacterium, Bdellovibrio phagocytosis characteristic may will die down, but the Bdellovibrio that the object of the invention is to filter out stability and high efficiency, so the present invention has its distinctive feature on the source of Bdellovibrio.
Before carrying out Bdellovibrio separation, the present invention needs to carry out specific pre-treatment to mud sample, to obtain Bdellovibrio stoste, the suspended particulate of the overwhelming majority, protozoon and virus can be removed by this kind of mode in centrifugal process, thus obtain the higher Bdellovibrio stoste of concentration, be conducive to sharp separation to Bdellovibrio, if do not carry out appropriate pre-treatment, can directly impact follow-up go out spot efficiency and Bdellovibrio separating effect.
In addition, Bdellovibrio is very high to the requirement of living environment, and in order to improve the propagation separation efficiency of Bdellovibrio further, the present invention is optimized again to the culture condition (proportioning, culture temperature etc. of pH, substratum and nutrient solution nutritive ingredient) in culturing process.If the pH in view of second pond active sludge is between 6.8 ~ 7.5, the present invention controls the scope of substratum and nutrient solution Optimal pH between 6.8 ~ 7.5; In addition, the growth needs Ca of Bdellovibrio is considered 2+, Mg 2+deng the absorption of trace element, substratum used in the present invention all additionally with the addition of the Ca of respective concentration 2+, Mg 2+, within the shortest time, obtain sufficient nutrition breed to be conducive to Bdellovibrio.Found that the speed in Bdellovibrio invasion host bacteria body is obviously accelerated, the spot time that goes out of Bdellovibrio also foreshortened to 2 ~ 3 days from now methodical 4 ~ 6 days, went out spot efficiency and significantly improved.
 
Utilize the Bdellovibrio that the present invention obtains, in its liquid multiplication culture process, speed and its own amplification speed of Bdellovibrio cracking host bacteria are obviously accelerated, and it still has the crack characteristic of efficient stable through Secondary Culture.But because Bdellovibrio individuality is small, the concentration after bacterial multiplication cannot be obtained with ascites, therefore weaken by what measure that Host Strains concentration in Host Strains and Bdellovibrio mixed solution reduces the bacterium liquid absorbancy caused the proliferate indirectly indicating Bdellovibrio.By research, Bdellovibrio proliferating liquid is obviously clear by turbid change at the 3rd day, and within the 4th day, absorbancy reduces to 0 substantially, and indicate that Host Strains is substantially cleaved and complete, Bdellovibrio growth concentration reaches the limit values.The efficient Bdellovibrio that the present invention is separated from second pond active sludge all has potential application prospect for the research of mud broken wall decrement and tail water process, has researching value.
Accompanying drawing explanation
Fig. 1 is the change curve of Bdellovibrio proliferating liquid with incubation time.
 
Embodiment
Now the invention will be further described with Figure of description in conjunction with the embodiments.
The material source that the present invention relates to is as follows:
1) mud sample takes from the active sludge of municipal sewage plant second pond.
2) in the present invention, the peptone yeast extract medium used, dilution nutrient broth medium, double-deck nutrient agar are on the basis of reference standard formula, carried out certain adjustment, and the pH controlling substratum is between 6.8 ~ 7.5, Optimal pH is 7.1.
The proportioning of peptone yeast extract medium is: peptone 9.5 ~ 10.5g/L, yeast extract paste 4.7 ~ 5.3g/L, sodium-chlor 9.5 ~ 10.5g/L, and agar powder mass concentration is 1.0% ~ 1.2%, ultrapure water 1000mL;
The proportioning of 1/500 dilution nutrient broth nutrient solution is: ultrapure water 1000mL, peptone 9.5 ~ 10.5g/L, extractum carnis 2.8 ~ 3.2g/L, sodium-chlor 4.7 ~ 5.3g/L, regulate pH after heating for dissolving between 6.8 ~ 7.5, again after high pressure steam sterilization 30min, by nutrient broth gradient dilution 500 times, the dilution nutrient broth nutrient solution of 1/500 can be obtained;
The proportioning of lower floor's nutrient agar is: in the ultrapure water of 1000mL, add 1/500 dilution nutrient broth nutrient solution 2 ~ 4mL, mass concentration be 0.25 ‰ ~ 0.35 ‰ CaCl 2, mass concentration be 0.40 ‰ ~ 0.50 ‰ MgCl 2be 1.0% ~ 1.2% agar powder with mass concentration;
The concrete proportioning of upper strata nutrient agar is: in the ultrapure water of 1000mL, add following material: 1/500 dilution nutrient broth nutrient solution 2 ~ 4mL, mass concentration are the CaCl of 0.25 ‰ ~ 0.35 ‰ 2, mass concentration be 0.40 ‰ ~ 0.50 ‰ MgCl 2, mass concentration is 0.5% ~ 0.6% agar powder;
Damping fluid is sterile phosphate buffer, regulates pH between 6.8 ~ 7.5;
Above-mentioned various substratum, nutrient solution and damping fluid all use high pressure steam sterilization 30min after preparation completes.
3) instrument used comprises biochemical aseptic operating platform, constant temperature water bath shaking table, constant temperature biochemical cultivation case, high-pressure steam sterilizing pan, pH meter, high speed freezing centrifuge, vortex oscillation instrument, 4 DEG C of refrigerators.
In order to reach the effect of high efficiency separation Bdellovibrio, the present invention proposes a kind of method of separation and purification Bdellovibrio from municipal sewage plant active sludge, the specific embodiments of the method is primarily of following 4 steps composition:
1. the screening of host bacteria and the preparation of bacteria suspension thereof
The screening process of host bacteria: first sample from the second pond active sludge of municipal sewage plant and preliminary treatment is carried out to mud, namely first get mud and carry out vortex oscillation 20 ~ 30min then standing 15 ~ 20min, then with sterile phosphate buffer, mud supernatant liquor carry out gradient dilution to 10 5~ 10 7doubly, then get each gradient dilution liquid 200 ~ 400 μ L respectively and coat on peptone yeast extract paste solid medium.After being coated with, place 10 ~ 20min, then moved in the constant incubator of 25 ~ 30 DEG C to be inverted and cultivate, optimum temps is 28 DEG C.After cultivation 18-24h, line is carried out for the isolated bacterium colony that flat board occurs be separated to obtain candidate's host bacteria, finally identify be separated candidate's host bacteria by gram staining method, therefrom screening and separating goes out the strong gram negative bacterium of stability as the host bacteria being separated Bdellovibrio.
The preparation flow of host bacteria suspension: join in nutrient broth nutrient solution by screening the Gram-negative host bacteria obtained, put into 25 ~ 30 DEG C, the constant temperature water bath shaking table of 150 ~ 200r/min cultivates 18 ~ 24h, then centrifugal 5 ~ 10min under 6000 ~ 9000r/min, get that precipitation sterile phosphate buffer is resuspended can obtain host bacteria suspension, and regulate its concentration 10 with sterile phosphate buffer 8~ 10 12cfu/mL(is bacterial concentration unit, is equivalent to individual/mL), preserve stand-by at being finally placed in 4 DEG C.
 
2. pair mud sample carries out pre-treatment to obtain Bdellovibrio stoste
Idiographic flow is: by the mud sample vortex oscillation 30 ~ 60min of screening Host Strains, after room temperature leaves standstill 20 ~ 30min, utilize high speed freezing centrifuge 4 ~ 10 DEG C, rotating speed be 800 × g ~ 1000 × g condition under centrifugal 15 ~ 25min, get supernatant liquor, then by supernatant liquor 4 ~ 10 DEG C, rotating speed be 15000 × g ~ 30000 × g under centrifugal 20 ~ 30min, get precipitation, and resuspended with sterile phosphate buffer, and the re-suspension liquid that obtains is Bdellovibrio stoste.
 
3. the isolation and purification of Bdellovibrio
First carry out the separation of Bdellovibrio according to following flow process, then at least repeat three separation and purification to the Bdellovibrio obtained according to following flow process, obtain pure Bdellovibrio, its concrete operations flow process is as follows:
(1) the double-deck agar plate method of dilution nutrient broth is adopted to carry out the separation of Bdellovibrio: first carry out gradient dilution to 10 to Bdellovibrio stoste 5~ 10 7doubly, get Bdellovibrio each gradient dilution liquid 500 ~ 700 μ L, join in the sterile test tube containing 500 ~ 700 μ L host bacteria suspension and mix, wherein the optimum concn of Host Strains is 10 10cfu/mL; And leave standstill 10 ~ 20min, then in test tube, add the top-layer agar substratum of 3 ~ 7mL insulation between 50 ~ 55 DEG C, after vortex oscillation mixing, pour into on the prefabricated bottom agar substratum solidified fast, rotate gently, make its uniform spreading on bottom agar substratum, leave standstill 10 ~ 20min, put into the constant temperature biochemical cultivation case cultivation 2 ~ 4d of 25 ~ 30 DEG C after top-layer agar culture medium solidifying after, top-layer agar substratum can grow Bdellovibrio bacterial plaque.。
(2) purifying of Bdellovibrio: first have picking on the substratum of Bdellovibrio bacterial plaque to isolate larger bacterial plaque from length and join in 10 ~ 20mL dilution nutrient broth nutrient solution, and add concentration 10 in nutrient solution 8~ 10 12host bacteria suspension 0.5 ~ 1.0mL between cfu/mL, after vortex oscillation mixing, be transferred to 25 ~ 30 DEG C, liquid increases 3 ~ 5d in the constant temperature water bath shaking table of 150 ~ 200r/min, the proliferating liquid that obtains is Bdellovibrio suspension; During this period, timing measure absorbancy, when proliferating liquid absorbancy drop to no longer change time, can illustrate that liquid has increased.Then by proliferating liquid gradient dilution to 10 5~ 10 7doubly, get each gradient dilution liquid and mix with host bacteria suspension.
After completing the separation of Bdellovibrio, according to above-mentioned steps 1), 2) flow process, to being separated at least re-treatment three times of the Bdellovibrio that obtains, to carry out the purifying of Bdellovibrio, obtain pure Bdellovibrio.Fig. 1 is Bdellovibrio absorbancy trend over time in liquid breeding, makes reference with sterilized water.
In the above separation and purification Bdellovibrio flow process of step 3, the concentration ratio that the Bdellovibrio gradient dilution liquid of each process mixes with host bacteria suspension is all 1/10 ~ 1/100.
4. the preparation of Bdellovibrio suspension
Pure Bdellovibrio embodiment step 3 obtained joins in Sterile dilution nutrient broth nutrient solution, and to add concentration be wherein 10 8~ 10 12the host bacteria suspension of cfu/mL, its volume accounts for 0.5% ~ 1% of dilution nutrient broth nutrient solution, and be transferred to 25 ~ 30 DEG C, cultivate 3 ~ 5d in the constant temperature water bath shaking table of 150 ~ 200r/min, the proliferating liquid obtained is pure Bdellovibrio suspension.
 
Above-described embodiment is only the preferred embodiment of the present invention; be noted that for those skilled in the art; under the premise without departing from the principles of the invention; some improvement and equivalent replacement can also be made; these improve the claims in the present invention and are equal to the technical scheme after replacing, and all fall into protection scope of the present invention.

Claims (5)

1. the method for separation and purification Bdellovibrio from active sludge, it is characterized in that, the method comprises the following steps:
1) preparation of screening and separating Host Strains and bacteria suspension thereof in mud sample: first using the active sludge of municipal sewage plant second pond as mud sample, therefrom separate Gram-negative bacteria, then using described Gram-negative bacteria as host bacteria, after liquid propagation 18 ~ 24h is carried out to it, obtain bacterial plaque through centrifugation; Finally the resuspended host bacteria suspension that obtains is carried out to obtained bacterial plaque sterile phosphate buffer, it is preserved at 4 DEG C;
2) pre-treatment is carried out to obtain Bdellovibrio stoste to mud sample, idiographic flow is: by the homology mud sample vortex oscillation 30 ~ 60min of screening Host Strains, after room temperature leaves standstill 20 ~ 30min, utilize high speed freezing centrifuge 4 ~ 10 DEG C, rotating speed be 800 × g ~ 1000 × g condition under centrifugal 15 ~ 25min, get supernatant liquor, then by supernatant liquor 4 ~ 10 DEG C, rotating speed be 15000 × g ~ 30000 × g under centrifugal 20 ~ 30min, get precipitation, and resuspended with sterile phosphate buffer, the re-suspension liquid that obtains is Bdellovibrio stoste;
3) separation of Bdellovibrio is carried out according to following flow process, then according to this flow process, three purifying are at least repeated to the mixed solution obtained, obtain pure Bdellovibrio: first adopt the double-deck agar plate method of dilution nutrient broth to carry out the separation of Bdellovibrio, then join in dilution nutrient broth nutrient solution by being separated the Bdellovibrio obtained, and add the host bacteria suspension that described step 1) obtains wherein, its volume accounts for 0.5% ~ 1% of dilution nutrient broth nutrient solution, after vortex oscillation mixing, be transferred to 25 ~ 30 DEG C, in the constant temperature water bath shaking table of 150 ~ 200r/min, liquid increases 3 ~ 5 days, by proliferating liquid gradient dilution 10 5~ 10 7doubly, get each gradient dilution liquid and mix with host bacteria suspension,
4) preparation of pure Bdellovibrio suspension: the bacterial plaque of the pure Bdellovibrio described step 3) obtained joins in Sterile dilution nutrient broth nutrient solution, and add the host bacteria suspension that described step 1) obtains wherein, its volume accounts for 0.5% ~ 1% of dilution nutrient broth nutrient solution, be transferred to 25 ~ 30 DEG C, cultivate 3 ~ 5 days in the constant temperature water bath shaking table of 150 ~ 200r/min, the proliferating liquid obtained is pure Bdellovibrio suspension.
2. the method for separation and purification Bdellovibrio from active sludge according to claim 1, it is characterized in that, the idiographic flow adopting the double-deck agar plate method of dilution nutrient broth to carry out the separation of Bdellovibrio in described step 3) is: carry out gradient dilution to Bdellovibrio stoste, get each gradient dilution liquid of Bdellovibrio and host bacteria suspension mixes, and add the top-layer agar substratum of 3 ~ 7mL insulation between 50 ~ 55 DEG C wherein, after vortex oscillation mixing, evenly be laid on the prefabricated bottom agar substratum solidified, the constant temperature biochemical cultivation case putting into 25 ~ 30 DEG C after top-layer agar culture medium solidifying is cultivated 2 ~ 4 days.
3. according to claim 1 or 2 from active sludge the method for separation and purification Bdellovibrio, it is characterized in that, in described step 3), the pH of the nutrient agar used in all separation and purification processes and dilution nutrient broth nutrient solution all needs to regulate between 6.8 ~ 7.5.
4. according to claim 1 or 2 from active sludge the method for separation and purification Bdellovibrio, it is characterized in that, in described step 3), in the nutrient agar used in double-deck agar plate method, also comprise the CaCl that mass concentration is 0.25 ‰ ~ 0.35 ‰ 2, mass concentration be 0.40 ‰ ~ 0.50 ‰ MgCl 2.
5. according to claim 1 or 2 from active sludge the method for separation and purification Bdellovibrio, it is characterized in that, the concentration of the host bacteria suspension used in described step 3) and step 4) is 10 8~ 10 12cfu/mL, in described step 3), the concentration ratio that Bdellovibrio gradient dilution liquid mixes with host bacteria suspension is 1/10 ~ 1/100.
CN201410587775.3A 2014-10-29 2014-10-29 Method for separating purified bdellovibrio from active sludge Pending CN104312963A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105254155A (en) * 2015-11-27 2016-01-20 余冉 Biological wall-breaking method for improving sludge dewatering performance
CN105969690A (en) * 2016-06-06 2016-09-28 东南大学 Phagocytosing type bacterium and application thereof in reducing sludge
CN108998386A (en) * 2018-07-09 2018-12-14 东南大学 A kind of phagocytosis type bacterium applied to deeply dehydrating sludge
CN109097298A (en) * 2018-08-08 2018-12-28 福建九为生物技术有限公司 A kind of method of enrichment culture method preparation phage bdellovibro preparation
CN110184222A (en) * 2019-06-06 2019-08-30 厦门惠盈动物科技有限公司 A kind of bacteriophagic Bdellovibrio freeze-dried powder preparation and its application

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LAURA HOBLEY: "Isolation and identification of Bdellovibrio and like organisms(BALOs) from various saltwater sites in the southern cape cod area,and analysis of their prey range specificity", 《MICROBIAL DIVERSITY》 *
何瑞阳等: "一株蛭弧菌的分离、纯化与PCR鉴定", 《安徽农学通报》 *
曾佳等: "淡水湖泊中类蛭弧菌的分离方法", 《矿物学报》 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105254155A (en) * 2015-11-27 2016-01-20 余冉 Biological wall-breaking method for improving sludge dewatering performance
CN105969690A (en) * 2016-06-06 2016-09-28 东南大学 Phagocytosing type bacterium and application thereof in reducing sludge
CN105969690B (en) * 2016-06-06 2019-10-11 东南大学 A kind of phagocytosis type bacterium and its application in mud decrement
CN108998386A (en) * 2018-07-09 2018-12-14 东南大学 A kind of phagocytosis type bacterium applied to deeply dehydrating sludge
CN108998386B (en) * 2018-07-09 2020-08-28 东南大学 Phagocytic bacteria applied to deep dehydration of sludge
CN109097298A (en) * 2018-08-08 2018-12-28 福建九为生物技术有限公司 A kind of method of enrichment culture method preparation phage bdellovibro preparation
CN109097298B (en) * 2018-08-08 2021-09-28 福建九为生物技术有限公司 Method for preparing bdellovibrio bacteriovorus preparation by enrichment culture method
CN110184222A (en) * 2019-06-06 2019-08-30 厦门惠盈动物科技有限公司 A kind of bacteriophagic Bdellovibrio freeze-dried powder preparation and its application

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Application publication date: 20150128