CN101455192A - Bdelloid rotifers mass culture method - Google Patents
Bdelloid rotifers mass culture method Download PDFInfo
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- CN101455192A CN101455192A CNA2009100448267A CN200910044826A CN101455192A CN 101455192 A CN101455192 A CN 101455192A CN A2009100448267 A CNA2009100448267 A CN A2009100448267A CN 200910044826 A CN200910044826 A CN 200910044826A CN 101455192 A CN101455192 A CN 101455192A
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Abstract
The invention relates to a large-scale culture method of bdelloid rotifers, and pertains to a novel bdelloid rotifers cultivation method and a novel bait process and use branch of technology.The present invention provides a bdelloid rotifers large-scale culture method by using non-living bait that is the wheat flour as bait,and is characterized in that the wheat flour after a biocidal treatment is added into in a certain proportion of sterilized water, the wheat flour and the sterilized water has a weight-bulk ratio of 0.75-0.85g, that is sterilized water of each 100 millilitre is added with wheat flour of 0.75-0.85g,and then the sterilized water added with wheat flour is performed with ultrasonication, stewing and subsiding for 24 hours, finally the supernatant fluid is took out and the PH value of the supernatant fluid is modulated to 5-7.The bait solution is then placed in the prepared culture solution of bdelloid rotifers, and is placed in a thermostat culture box of 25 DEG C for 12 days, as a result, density of the bdelloid rotifers is up to 100-200ind.Bait source of the invention is abroad, the manufacturing is convenient, the cultivation course can be simplified and the cost can be reduced.
Description
Technical field
The present invention relates to a large amount of cultured method of a kind of Bdelloid rotifers, belong to the new cultural method of Bdelloid rotifers and processing of novel bait and operation technique field.
Background technology
Bdelloid rotifers (bdelloid rotifer) extensively is present in and it is characterized by the forked toe of tail end in water purification and the sewerage ecological environment, but the body of gland secreting mucus in the toe makes its surface that can adhere to other solid (activated sludge, biomembrane) with battalion's epiphytism.Because of its ability with the strong suspended solid of ingesting is subjected to paying close attention to widely.Common Bdelloid rotifers has spinning roller to belong to (Philodina), wheel Eimeria (Rotaria) and place wheel genus (Habrotrocha) wheel animalcule in bioreactor for disposing polluted water.Bdelloid rotifers not only can reach purifying water body and reduce the effect of surplus sludge volume by the suspended solid of ingesting, and the activity to activated sludge or biomembranous formation, bacterium also has facilitation simultaneously.At present a lot of to the research of wheel animalcule cultural method, but mainly with arm tail wheel Eimeria (Brachinorus), crystal sac wheel Eimeria (Asplanchna), it is main that vertebra wheel Eimeria (Notommata) waits some biotype wheel animalcules that swim.And it is few in number to the research of Bdelloid rotifers that important function is arranged in the Sewage Biological Treatment.
Tradition is used to cultivate the bait of Bdelloid rotifers based on biological feed bacterium and algae.Though bacterium and algae can be satisfied the needs that wheel animalcule preys on, the bacterial culture cycle is long, and bacterial solution needs fresh configuration to use inconvenience simultaneously, also very easily flocculates in the cultivation and should not prey on.Though and algae nutrition is very high, usually particle diameter is excessive is not suitable for the Bdelloid rotifers predation, and algae also needs the certain culture condition as organism simultaneously, therefore uses the not too convenient expense of cultivating that improved simultaneously.
Summary of the invention
The object of the invention provides that a kind of abiotic bait of using---wheat flour is cultivated the method for Bdelloid rotifers in a large number.
The present invention is a large amount of cultured method of a kind of Bdelloid rotifers, it is characterized in that having following step and feature:
A, the food that utilizes abiotic bait solution to grow as Bdelloid rotifers.Its preparation process is as follows: get a certain amount of wheat flour as the bait raw material, at first with the wheat flour sterilization treatment, add a certain amount of sterile water again, the w/v of wheat flour and sterile water is 0.75～0.85g/100mL, promptly adds the wheat flour of 0.75～0.85g in the distilled water of every 100mL; The solution that configures again through ultrasonic disruption, is wanted suitable cooling in the ultrasonic disruption process, make wheat flour solution in the pasty state in order to avoid produce a large amount of heats in the shattering process; Flour soln sedimentation after the fragmentation is discarded the flour of bottom precipitation after 1 hour, removing the flour particle of big particle diameter, and guarantee its epigranular; Get its supernatant behind the sedimentation 24h once more, the bait solution that finally obtains is that the concentration of flour suspension is 5 * 10
6About grain/mL, and particle diameter is distributed in 0.2～0.3 mu m range; PH value with culture fluid is adjusted in 5～7 scopes at last; Be stored in the refrigerator bait solution for preparing stand-by.
B, the above-mentioned abiotic bait solution that makes is incorporated in the aseptic culture dish of preprepared, adds the described bait solution of 10～100 μ L in every milliliter of culture fluid.
C, inoculate Bdelloid rotifers in above-mentioned culture fluid, the initial density of inoculation is 1ind/mL.Be placed in 25 ℃ of constant incubators and cultivate, cultivate after 12 days, the density of wheel animalcule can reach 100～200ind/mL in the culture dish.
Advantage of the present invention and characteristics are as described below:
Characteristics of the present invention are to adopt the wheat flour that cheaply is easy to get as the bait of cultivating Bdelloid rotifers, the manufacturing process simple possible of bait, and the bait of making is easy to use, not perishable, be easy to preserve, and can reduce the cultivation cost.And contain a large amount of mineral elements in the common wheat flour, and amino acid, vitamin and kinds of specific polysaccharides can provide prescribing adequate nutrition can satisfy the various needs of wheel animalcule growth.Through cultivating, the wheel animalcule density in the culture dish can reach 100-200ind/mL.
In addition, experimental study draws, and the grow pH value scope of optimal flour soln of Bdelloid rotifers is 5～7, and the pH value that growth is best is 6.With this understanding, the wheel animalcule healthy growth, breeding is fast.
Description of drawings
The population density change curve of wheel animalcule when Fig. 1 is bait for tradition with mixed cell.
Fig. 2 is the population density change curve with flour soln of the present invention wheel animalcule when cultivating bait.
Specific growth rate size comparison diagram when Fig. 3 cultivates wheel animalcule for flour soln of the present invention is used to cultivate wheel animalcule and traditional mixed cell.
Embodiment
After now specific embodiments of the invention being described in.
Embodiment 1
Bdelloid rotifers incubation and step in the present embodiment are as follows:
(1) take by weighing wheat flour 3.8 gram, be incorporated in 500 milliliters the sterile water through sterilization treatment, then with this solution through ultrasonic disruption, suitably cooling makes wheat flour solution in the pasty state in order to avoid produce a large amount of heats in ultrasonic disruption; Flour soln sedimentation after the fragmentation is discarded the flour of bottom precipitation after 1 hour, removing the flour particle of big particle diameter, and guarantee its epigranular; Get its supernatant behind the sedimentation 24h once more, the bait solution that finally obtains is that the concentration of flour suspension is 5 * 10
6About grain/mL, and particle diameter is distributed in 0.2～0.3 mu m range; PH value with culture fluid is adjusted in 5～7 scopes at last; Be stored in the refrigerator bait solution for preparing stand-by.
(2) the above-mentioned abiotic bait solution that makes is incorporated in the aseptic culture dish of preprepared, adds the different described bait solution of measuring in every milliliter of culture fluid.The three groups of parallel samples of bait solution composition that promptly add 10,50,100 microlitres respectively.
(3) place 25 ℃ of constant incubators to cultivate respectively on above-mentioned three groups of samples.After cultivating in 12 days, the averag density of wheel animalcule can reach 102.7ind/mL.
The contrast experiment
Fig. 1, Fig. 2 and shown in Figure 3 in contrast and experiment with flour soln of the present invention and conventional hybrid bacterium during such as the accompanying drawing as the bait of cultivating wheel animalcule.
The population density variation diagram of wheel animalcule when Fig. 1 represents to be bait with the mixed cell, by can drawing among the figure, it is 34.33ind/mL that wheel animalcule can reach maximum population density when being the cultivation of bait with the mixed cell.The population density variation diagram of wheel animalcule when Fig. 2 represents that flour soln of the present invention is used to cultivate, by can drawing among the figure, wheel animalcule is that can to reach maximum population density under the cultivation of bait be 102.7ind/mL with flour soln of the present invention.The specific growth rate size comparison diagram of wheel animalcule can be drawn by figure when Fig. 3 represented that flour soln of the present invention is used to cultivate wheel animalcule and is bait with the mixed cell, was that the specific growth rate size of wheel animalcule is 0.6109d under the cultivation of bait with flour soln of the present invention
-1, be under the cultivation of bait with the mixed cell, the specific growth rate of wheel animalcule size is 0.3231d
-1
By Fig. 1,2,3 relatively get, bait solution of the present invention extremely helps the growth of Bdelloid rotifers population, under it was cultivated, it was situation under bait is cultivated that population density of wheel animalcule and specific growth rate are far longer than with the mixed cell, as seen, the present invention is not only feasible as the bait of Bdelloid rotifers, and has culture effect preferably.
Claims (1)
1, a large amount of cultured method of a kind of Bdelloid rotifers is characterized in that having following step and feature:
A, the food that utilizes abiotic bait solution to grow as Bdelloid rotifers; Its preparation process is as follows: get a certain amount of wheat flour as the bait raw material, at first with the wheat flour sterilization treatment, add a certain amount of sterile water again, the w/v of wheat flour and sterile water is 0.75～0.85g/100mL, promptly adds the wheat flour of 0.75～0.85g in the distilled water of every 100mL; The solution that configures again through ultrasonic disruption, is wanted suitable cooling in the ultrasonic disruption process, make wheat flour solution in the pasty state in order to avoid produce a large amount of heats in the shattering process; Flour soln sedimentation after the fragmentation is discarded the flour of bottom precipitation after 1 hour, removing the flour particle of big particle diameter, and guarantee its epigranular; Get its supernatant behind the sedimentation 24h once more, the bait solution that finally obtains is that the concentration of flour suspension is 5 * 10
6About grain/mL, and particle diameter is distributed in 0.2～0.3 mu m range; PH value with culture fluid is adjusted in 5～7 scopes at last; Be stored in the refrigerator bait solution for preparing stand-by;
B, the above-mentioned abiotic bait solution that makes is incorporated in the aseptic culture dish of preprepared, adds the described bait solution of 10～100 μ L in every milliliter of culture fluid;
C, inoculate Bdelloid rotifers in above-mentioned culture fluid, the initial density of inoculation is 1ind/mL; Be placed in 25 ℃ of constant incubators and cultivate, cultivate after 12 days, the density of wheel animalcule can reach 100～200ind/mL in the culture dish.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009100448267A CN101455192A (en) | 2009-01-04 | 2009-01-04 | Bdelloid rotifers mass culture method |
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| CNA2009100448267A CN101455192A (en) | 2009-01-04 | 2009-01-04 | Bdelloid rotifers mass culture method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103053476A (en) * | 2012-12-05 | 2013-04-24 | 大连海洋大学 | Cultivating method for obligate parthenogenesis rotifera population |
| CN105647831A (en) * | 2016-03-03 | 2016-06-08 | 上海大学 | Method for accelerating growth of flocculating bacteria by using Philodina culture liquid |
| CN105779345A (en) * | 2016-03-29 | 2016-07-20 | 上海大学 | Method for promoting growth of zymomonas mobilis by using philodina culture solution |
| CN113816498A (en) * | 2021-10-29 | 2021-12-21 | 上海大学 | Method for improving biological treatment efficiency of wastewater by adding hirudo |
-
2009
- 2009-01-04 CN CNA2009100448267A patent/CN101455192A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103053476A (en) * | 2012-12-05 | 2013-04-24 | 大连海洋大学 | Cultivating method for obligate parthenogenesis rotifera population |
| CN103053476B (en) * | 2012-12-05 | 2015-07-22 | 大连海洋大学 | Cultivating method for obligate parthenogenesis rotifera population |
| CN105647831A (en) * | 2016-03-03 | 2016-06-08 | 上海大学 | Method for accelerating growth of flocculating bacteria by using Philodina culture liquid |
| CN105647831B (en) * | 2016-03-03 | 2019-06-25 | 上海大学 | Promote the method for flocculability bacterial growth with spinning roller worm culture solution |
| 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 |
| CN113816498A (en) * | 2021-10-29 | 2021-12-21 | 上海大学 | Method for improving biological treatment efficiency of wastewater by adding hirudo |
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Open date: 20090617 |