CN104568563B - A kind of device and method of soil nematodes separation and Extraction - Google Patents
A kind of device and method of soil nematodes separation and Extraction Download PDFInfo
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- CN104568563B CN104568563B CN201510019727.9A CN201510019727A CN104568563B CN 104568563 B CN104568563 B CN 104568563B CN 201510019727 A CN201510019727 A CN 201510019727A CN 104568563 B CN104568563 B CN 104568563B
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Abstract
The present invention discloses a kind of device and method of soil nematodes separation and Extraction, and device includes support, soil cup, agitator, bolter, water injection system, collection vessel, and agitator is placed in soil cup, and soil cup bottom has a liquid outlet；Bolter includes the first order and second level bolter, and in the second level, the lower end of bolter, which is provided with, draws liquid mouth；The first order, second level bolter are respectively 40 mesh and 400 mesh；Water injection system includes sparge pipe, and sparge pipe is arranged at sieve wall.The present invention is separated using two-stage, and uses special water spray tubular construction so that nematode can dramatically be separated, and not omit.
Description
Technical field
The present invention relates to a kind of separation of soil nematodes and the device and method obtained.
Background technology
Soil nematodes is as the second largest invertebrate monoid that insect is only second in nature, because it has distribution wide
It is general, occupy in food chain key position, be rapid to the nutrient and disturbance response of environment, Structural Identification method is simple and structure with
There is the features such as obvious correlation between function, be often counted as indicating one of important indicator of soil health.It is fast from soil
Speed and comprehensively nematode is separated be study nematode the first step, at present nematologist typically with funnel method, screening eluriate
Method, Centrifugal Flotation method and tray method separate nematode from soil.
1st, funnel method and tray method
Funnel method is a kind of conventional method, and major technique is that one section of rubber tube and rubber tube rear end use are connect in funnel end
Spring clip clamps.In use, placing one layer of screen cloth first in funnel, two layers of gauze and a layer line worm filter paper are placed thereon.Soil
After sample is uniformly layered on filter paper, submergence soil is added water to.Respectively after 24,36,48h, clip is opened, is released in rubber tube
Water in small beaker.Directly the water of acquisition is counted and differentiated.This method is to utilize the hydrotaxis of nematode and itself
Weight, depart from soil successively.The advantages of this method is simple and cheap.Shortcoming is that the nematode obtained is confined to through filter paper hole
It is the strong nematode of energy that gap, which enters in water, in the case of more than soil sample, can influence to obtain because the activity of nematode is low
The efficiency of nematode；In addition, the time of extraction is generally 48 hours.Time is short to be difficult to catch mobile slow nematode；Furthermore if behaviour
It is improper to make, and nematode liquid can be caused muddy.
The stainless steel screen tray of 10 mesh, i.e., be put into supporting tray, put on screen tray by the tray method that the country uses at present
Put two layers of gauze and a layer line worm filter paper.General principle is close with funnel method, and only soil sample paves the area on filter paper
It is larger, be advantageous to the activity of nematode.
2nd, sieve method is eluriated
Sieve method is eluriated mainly to be separated using the difference of difference in size between nematode and soil constituent and proportion.
This method mainly will add water in soil, after screening from coarse to fine several times, most of nematode rests on carefully for suspension part
On sieve.This method does not need special installation, and method simply waits also without prolonged.In recent years, scholar surrounds nematode
Many improvement have been carried out from the separation of soil particle, have such as focused on the gradient precipitation of separation of sediment.But in separation process, if not
It is careful then have many nematodes and stay in soil；The nematode liquid liquid sieved in addition influences because causing muddiness containing some silts
Observation to nematode.
3rd, sucrose solution Centrifugal Flotation method
Centrifugal Flotation method Main Basiss are that the proportion of nematode is about 1.08g/m3, it is more bigger than the proportion of water, in soil supension
Middle nematode mixes with dirt bed.If with the slightly larger sucrose solution (1.18g/m of proportion3), it is possible to nematode from suspension
In separate.Have main steps that and add water to be placed in centrifuge tube soil sample, after centrifuging a few minutes, abandoning supernatant.Then add
800gL-1 sucrose solutions stir evenly, and centrifuge a few minutes again, by supernatant note into different size sieve (300 mesh, 400 mesh,
500 mesh), and rinsed with water.After obtaining the nematode in screen cloth, counted and classified.This method is relatively easy and rapid.
The stirring link being related in operating process can cause soil to disperse because centrifugation tube space is limited；In addition after first centrifugation
The very consolidation of the soil particle containing nematode, nematode can be influenceed from the further separation in soil, especially heavy texture
Soil is then even more serious.Therefore, although the method is quick, due in soil dispersion separation process, if do not had carefully perhaps
More nematodes are stayed in soil, and screening liquid is easily muddy, is unfavorable for the observation of nematode.
To solve the problems, such as the sucrose Centrifugal Flotation method complex steps of existing soil nematodes.There is scholar to invent and use high concentration cl
Change potassium solution flotation nematode.Main process is to add soil in Klorvess Liquid, after stirring, then stands to obtain supernatant and precipitation
Thing, then replace pedotheque to repeat aforesaid operations sediment, injected after obtained supernatant is merged in mesh screen, then use water
Backwash mesh screen.This method avoid the centrifuging process in sucrose extraction process, but still need to repeatedly pedotheque or precipitation
Thing stirring, stand.The supernatant being collected into rinses mesh screen with water and obtains nematode repeatedly.Resulting supernatant is muddy, sediment
Nematode collect not congruent problem.
Because soil nematodes is inhabited in the fine capillaries formed between soil clay particle, it is possible to because clay obtains not
To fully dispersed and can not dissociate and deal.So in nematode extracting methods numerous at present, some is not stirred soil
Mix, have it is main be stirred with glass bar, these are all unable to reach to the fully dispersed of soil particle.
In addition, it is necessary to from being carried out with microscope after being separated to soil nematodes from soil, it is currently used be exactly containing
The aqueous solution of nematode.The nematode liquid that method obtains at present substantially has two kinds of situations：A kind of is that can obtain cleaner nematode liquid, but
In cleaner solution is obtained, it is possible to cause a large amount of nematode abandoneds in the transfer process of sediment, has a strong impact on and grind
The effect studied carefully.Although another situation is exactly to obtain the nematode liquid of complete number, more soil can be contained in nematode liquid
Particle.This is two conflicting aspects.
Sum it up, common problem existing for above-mentioned each method is exactly, first, nematode can not thoroughly be divided from soil
Separate out and, second, the nematode separated can not be fully collected.The aspect of the above two all causes soil nematodes statistical number
The inaccuracy of amount, the accuracy of soil investigation can be influenceed.
The content of invention
In order to which the nematode in soil is separated to greatest extent, and the soil nematodes liquid of clarification is obtained, reduced
The workload for obtaining nematode is rinsed, the present invention provides dress and the side of a kind of convenient, fast and efficient soil nematodes separation and Extraction
Method.Using the device and corresponding separating and extracting process, limits can be use up nematode is extracted from soil, it is ensured that carry
The accuracy of access amount, believable guarantee is provided for agrology research.
One aspect of the present invention provides a kind of soil nematodes separating extraction device, and technical scheme is：Including support, soil cup,
Agitator, bolter, water injection system, collection vessel；The soil cup be a bottom at least have a liquid outlet cup, institute
State and a mozzle is connected on liquid outlet, a plug valve is set on mozzle；The agitator is placed in the soil cup；Described point
Include first order bolter and second level bolter from sieve, first order bolter is horizontally or diagonally places, second level bolter
It is then slant setting, in the second level, the lower end of bolter, which is provided with, draws liquid mouth；First order bolter is 40 mesh, and the second level separates
Sieve as 400 mesh；Bolter include sieve bottom and sieve wall two parts, sieve wall be along sieve feather edge naturally upwards linking form, sieve bottom and
The place for sieving wall linking forms circular arc transition；The water injection system includes sparge pipe, the water outlet of the sparge pipe along sieve wall to
The lower end of sieve is washed away.
Preferably, the soil cup is the tapered cup in a bottom, and the liquid outlet is arranged at tapered bottom.
Preferably, the agitator includes rotary shaft and stirring wheel, and stirring wheel is radially fixed in ring week by several
Blade composition in rotary shaft, every blade are tilted with the horizontal certain angle, and plane body or spiral are shaped as per blade
Shape curved body.
A kind of preferred scheme is：The sieve cornice has water fender, and the water fender is horizontally sieved along upper by sieve wall
Sub- inner side is rolled over, and is then rolled over and is formed straight down again, and the water fender is in sieve one whole circle of inboard formation；The now water spray
Pipe includes top sparge pipe, and the top sparge pipe is tied up into a whole circle in the region between the water fender and sieve wall, described
Carry apopore on the sparge pipe of top, water outlet along on the inside of sieve wall to flowing down.
Another preferred scheme is：The sieve cornice has a water fender, the water fender be by sieve wall along it is upper horizontally
Rolled on the inside of sieve, then roll over and formed straight down again, the water fender is in the inboard formation half-turn of sieve；The sparge pipe bag
Top sparge pipe is included, is tied up corresponding to the top sparge pipe into half-turn in the region between the water fender and sieve wall, it is described
Carry apopore on the sparge pipe of top, water outlet along on the inside of sieve wall to flowing down；The water fender and the top sparge pipe all positions
In the high-end position of inclined bolter；The sparge pipe also includes bottom sparge pipe, and the bottom sparge pipe is tied up in institute into whole circle
The bottom of sieve wall is stated, carry apopore on the bottom sparge pipe, water outlet direction is towards sieve inner side.
Further, the top sparge pipe is tied up on the sieve wall, or is tied up on the water fender.
The water spray bore is 5mm, dispersed on pipe to be provided with several apopores, is further opened with pipe
One water inlet, the water inlet are located at the top of inclined sieve, connect outside the water inlet and add wetting system.
The top sparge pipe is bundled on distance sieve wall along down at 1cm, and height is less than the water fender herein
Highly；The bottom sparge pipe is bundled in distance sieve bottom up at 1cm.
In the case that the first order bolter and second level bolter are all slant setting, incline direction is on the contrary, and institute
State second level bolter angle of inclination and be more than the first order bolter.
Another aspect of the present invention provides a kind of method of soil nematodes separation and Extraction, comprises the following steps：
1) macroscopic debris is removed to carrying out simple process for trying fresh soil first；
2) being put into for trying fresh soil in soil cup for 100g is weighed, 400ml distilled water or running water is added, starts stirring
Device, with 50~100 revs/min of speed, stir 5 minutes；
3) plug valve on soil cup is opened, the collecting pipe below sieve is opened, goes out the native liquid stream in soil cup, pass through
Following first order bolter and second level bolter, flowed out from collecting pipe, into collection vessel；
4) when native liquid is run up to above the bolter of the second level, it is gently mixed with glass bar, while opens and add wetting system, to
Sparge pipe adds water, water is washed away sieve wall and sieve bottom, native liquid is flowed to liquid mouth direction is drawn；
5) residual of native liquid in soil cup is checked, is rinsed with a small amount of water, until being remained in soil cup without native liquid, the first fraction
Untill native liquid clarification under being sieved from sieve, plug valve is then shut off；
6) constantly with wetting system is added, by adding water to sparge pipe, flushing is accumulated in the native liquid above the bolter of the second level；See
The native liquid through second level bolter is examined, untill through the clarification of the native liquid of bolter；
7) collection vessel is collected into nematode suspension, by precipitation, is transferred to after removing supernatant liquor in centrifuge tube, collection
Container is rinsed well with water, and suspension is centrifuged, and 2000~3000 rev/min, then removes supernatant again；
8) 800g/L sucrose solutions are added in centrifuge tube, are centrifuged 2 minutes under 2000 revs/min of speed, by supernatant
The sieve noted into 500 mesh is sieved, and can obtain clean nematode with water flushing sieve.
Due to take above technical scheme and make the invention has the advantages that：
1st, realize fully dispersed to soil particle.The device is contacted using inclined blade with native liquid, is had and soil
Contact area is big, and during wheel rotation, native liquid by blade, can further increase the dynamics of stirring.
2nd, the speed that can control native liquid stream to go out.Native liquid enters bolter by cock, if excessive velocities, with that may cause
Native liquid rotary control knob, can not can open or close piston at any time too soon in the excessive of bolter.
3rd, repeatedly stirring can be achieved., can be according to soil property to be sufficiently scattered because the native liquid precipitate speed being dispersed with stirring is fast
Needs and repeatedly stirred.
4th, nematode loss is reduced.Soil cup portions are arcuate structure, are on the one hand reduce impeller and soil cup bottom direct
Contact and injure nematode, be also beneficial in addition to remnants native liquid clean, avoid because native liquid remaining and caused by nematode population
Reduction, so as to improve obtain nematode efficiency.
5th, bolter is easy to rinse.Sparge pipe is designed on sieve wall, wall and sieve bottom can be sieved with auto-flushing.In addition, water column
From different directions, a water filling, sieve wall and sieve bottom can be rinsed from all directions, reduce the work to be fed water during flushing
Measure, also reduce the pressure to nematode, be advantageous to the maintenance of nematode vigor.
6th, the acquisition of nematode is advantageous to.Inclined design is carried out to 400 mesh bolters, may be such that the water for rinsing nematode flows naturally
It is collected at the collecting pipe of lower end, and then to nematode aqueous suspensions.The grogs that the nematode suspension that collecting pipe is collected into also has is equal
More than 0.038mm, the native liquid for obtaining nematode is further precipitated and can be shortened with sucrose solution flotation, time, reduces nematode
Dehydration in sucrose solution, and the nematode solution clarification obtained, are easy to further collect and analyze.
Brief description of the drawings
Fig. 1 is a kind of overall structure diagram of embodiment of device；
Fig. 2 is a kind of example structure schematic diagram of soil cup；
Fig. 3 is a kind of example structure schematic diagram of agitator；
Fig. 4 is the structural representation of bolter (without water fender)；
Fig. 5 is that bolter combines embodiment elevational schematic with one kind of sparge pipe；
Fig. 6-1 is that bolter combines embodiment elevational schematic with the another kind of sparge pipe；
Fig. 6-2 is that bolter combines embodiment schematic top plan view with the another kind of sparge pipe；
Fig. 7 is the schematic diagram of sparge pipe.
Embodiment
The present invention is described in detail with reference to the accompanying drawings and examples.But those skilled in the art should know
Road, following examples are not the unique restriction made to technical solution of the present invention, every in technical solution of the present invention spirit and real
Any equivalents done under matter or change, it is regarded as belonging to protection scope of the present invention.
Content of the present invention mainly has two aspects, first, nematode is obtained with being separated in soil to greatest extent
To the cleaner soil supension containing nematode, this process mainly includes：
1) soil nematodes is free：Mainly to adding water to stir for examination soil sample, nematode is set to be separated out from soil capillary water middle reaches
Come；
2) screen out big particle and small soil powder glues particulate：Mainly the native liquid by disperseing is sieved,
Removal be more than 40 mesh (0.42mm) coarse sand granule and the silty sand and clay less than 400 mesh (0.038mm), and be collected into containing compared with
The nematode suspension of fine sand.
Second, nematode is separated from suspension, the nematode solution of clarification is obtained, to obtain accurate nematode population.For
Said process is completed, the corollary apparatus and corresponding method that the present invention uses are described as follows：
One, constructional device
As shown in figure 1, the device that the present invention takes mainly is included with lower component：Support, soil cup 1, agitator 2, separation
Sieve 3,4, water injection system, collecting pipe 7, collection vessel 8.To the part and application method of the present apparatus, said below in conjunction with the accompanying drawings
It is bright：
Soil cup 1
As shown in Fig. 2 soil cup 1 is the cup that a bottom has a liquid outlet 11 at least.For the shape of soil cup
Strict limitation is not done, but several preferred embodiments can be provided：First, soil cup is the flat cup of a cylinder, at cup bottom
Several liquid outlets are issued in portion；Second, soil cup is a taper or the cup for being similar to taper, can issue one in cone bottom goes out liquid
Mouthful；Third, it is cylinder that soil cup, which is a top, bottom is hemispheric cup-shaped (also referred to as bowl-shape), is opened in bowl bottom
Has a liquid outlet；Fourth, it is cylinder that soil cup, which is a top, bottom is the cup of taper, and a liquid outlet is issued in cone bottom.
In several embodiments, flat cup is inconvenient, first, poor fluidity is, it is necessary to set several liquid outlets to be just easy to different positions
The liquid outflow put, operation are cumbersome；Second, stirring is inconvenient, flat bottoms liquid is not easy to be stirred to delay is serious.For
Shape bottom of a cup is shunk in spherical or taper etc., opens up a liquid outlet can, and bottom of cups amount retained is small, is easily stirred,
So such cup is suitable for selection.Selected depth of the present invention and the larger soil cup of span (diameter 15cm, height
10cm), and without using elongated centrifuge tube, more sample soil can be loaded, also allow for being put into agitator.Ensureing to stir volume
While sufficiently large, it can also be ensured that stirring thing not by the viscous stop of wall of cup, make stirring thing along its wall of cup under Gravitative Loads from
So flow down, curved surface can be advantageous to stirring thing to be assembled to bottom of cup center, and this cup-shaped is applied to being dispersed with stirring for soil thing.
The liquid outlet size of control bottom of cups can control out the flow of liquid, for the specificity of soil nematodes,
The diameter design of liquid outlet 11, can be to allow nematode to flow out in 2.0cm or so.In the outer bottom of soil cup, a mozzle is set
12, it is connected on liquid outlet 11, liquid outlet 11 is communicated with mozzle 12, and mozzle can be welded on liquid outlet, can also
It is to be connected to sealing flange on liquid outlet.A plug valve 13 is crossed on mozzle 12.Due to having consent on plug valve, so can
With the anglec of rotation by controlling plug valve, to control the aperture of consent and mozzle, aperture is bigger, and liquid outlet quantity is bigger, aperture
Smaller, liquid outlet quantity is smaller.Plug valve belongs to common part.Such soil cup structure, it is to be made according to separatory funnel principle, therefore
The advantages of with separatory funnel.
Agitator 2
Agitator 2 is placed in soil cup, can be hand mixer, or electric mixer.A kind of hand operated mixing
The embodiment of device device is：The hand mixer includes handwheel 21, rotary shaft 22 and stirring wheel 23, as shown in figure 3, handwheel 21 connects
At the top of rotary shaft 22, stirring wheel 23 is connected to the bottom of rotary shaft 22, and rotary shaft 22 and stirring wheel 23 extend into soil cup
The inside, stirring wheel reach middle and lower part position in cup.The diameter of rotary shaft 22 about 2cm, high about 15cm.Stirring wheel 23 can be several and stir
Mixing rod, radially ring week is fixed in rotary shaft 22 (as shown in Figure 2)；Preferably can also be to be made by several blades, preferably
, every blade with the horizontal certain angle tilt, preferably about 30~45 °, per blade shape may be selected plane body,
Also helical form curved body may be selected, length and width, thickness ratio are about 16:10:3, these are not shown in illustration is implemented.Select leaf
Chip stirring wheel, agitation contact area can be made to be more than stirring rod；Blade lean, or blade selection helicoid, can reduce
Resistance is stirred, strengthens stirring dynamics.Stirring rod or blade preferably selection sheet rubber or plastic sheet with certain toughness are made.
In order that agitator rests on certain altitude in soil cup, the mode taken is that rotary shaft 22 is supported on into a support
On part, the support member can be a support, be preferably a cup lid 24.Support and cup lid are supported on soil cup rim of a cup,
Cup lid is then with more protection soil cup, from the effect of liquid splash in cup.The mode that rotary shaft 22 is supported on support member can
To there is two kinds：One, rotary shaft 22 can be supported on support or cup lid (as shown in figures 1 and 3) by the way of reducing, propped up
Section diameter more than frame or cup lid is big, the section diameter below support or cup lid is small be placed in agitator it is same in soil cup
When, support or cup lid are buckled in soil cup rim of a cup, play a part of support to rotary shaft, and rotary shaft is stuck on support or cup lid, and
Soil cup bottom of a cup can not be fallen into；Two, rotary shaft 22 can use the part for exposing support member on the rotary shaft is horizontal to insert a thin rod
Mode be supported on support or cup lid (do not provide this implement illustration).It is that activity wears relation between rotary shaft and support member,
Support member is motionless when rotary shaft rotates, and a bearing can be set by being worn on support member at the hole of rotary shaft.
Agitator 2 can also be the agitator of motor driven, on the basis of the hand mixer of foregoing description, now only
Handwheel 21 need to be cancelled, connect a motor in the upper end of rotary shaft 22, this belongs to routine techniques, and therefore not to repeat here, motor control
System can more control the speed of stirring very well.
Agitator is placed in soil cup, soil sample and a certain amount of water is added in soil cup, by line by the way of stirring
Worm soil is stirred scattered, and nematode is separated from soil to greatest extent, completes the free task of soil nematodes.Connect
Get off and exactly soil particle larger in soil supension and the viscous small package of less powder are gone out, leave behind outstanding containing nematode
Liquid.
The method of screening is to set two-stage bolter in the lower section of soil cup, and first order bolter 3 is undertaken on soil cup
Lower section, second level bolter 4 are undertaken on the lower section of first order bolter 3.In the present invention, first order bolter 3 to be horizontal or
Slant setting can, second level bolter 4 is then slant setting；And if this two-stage bolter is all tilted-putted premise
Under, then preferably two sieves are opposite direction slant setting, the place that first order screening thing falls is exactly the
The high-end position of two level sieve, first order screenings can so be slipped in by second level screening process along the high-end position of sieve
The low side position of sieve, so as to increase sieving time and sieving mileage, so as to improve screening effect.
And when the first order 3 slant setting of bolter, it is smaller than the angle of inclination of second level bolter 4 preferably, this
Sample can enable first order bolter smoothly catch the soil supension that soil cup flows down, and soil and nematode are gathered in substantially
The middle section sieving of sieve, and will not gather and be detained in the corner of sieve；And the angle of inclination of second level bolter is slightly
It is big, it the oversize after the separation of the second level is successfully flowed in collection vessel.Usually, we select first
Level bolter is that 10 °~15 ° angles of inclination are placed, and second level bolter is that 15 °~20 ° angles of inclination are placed.
The size of soil nematodes is typically in 0.04mm~0.4mm scopes, so we can set two-stage bolter respectively
It is set to 40 mesh sieves (0.42mm) and 400 mesh sieves (0.038mm), 40 mesh of first order bolter, 400 mesh of second level bolter,
The coarse sand granule that so can both make more than nematode is stored in above first order bolter, it is not necessary to is participated in the second-order separation, can be made again
Silty sand and clay less than nematode are fallen within below the bolter of the second level, without entering collection vessel with nematode suspension, thus
The particulate matter in nematode suspension in addition to nematode size can be made to reduce as far as possible, be advantageous to the further extraction of nematode.Top
Bolter can be smaller with diameter, and the bolter of lower section can be larger, and can so catch as far as possible by first order bolter
The object fallen.
The structure of two-stage bolter has points of resemblance, as shown in figure 4, bolter includes sieve bottom 41 and sieve two parts of wall 42,
Sieve bottom 41 is made by screen cloth, is divided into two kinds of meshes of 40 mesh and 400 mesh by rank；Sieve wall be along sieve feather edge naturally upwards be connected and
Into sieve wall 42 does not have mesh, and sieving wall and sieve bottom can all be made up of stainless-steel sheet.Formed between the place that sieve bottom is connected with sieving wall
Circular arc transition；And the position being preferably connected in sieve wall with sieving bottom, by the way of overlapping connection, in case sieve bottom takes off with sieve wall
Fall.Connection uses circular arc transient mode between sieve bottom and sieve wall, can avoid the generation at dead angle, so when nematode suspension is from sieve
When wall flows down, it can be flow to along smooth arc-shaped surface in the sieve big plane in bottom, the delay of nematode and particulate matter will not be caused.Phase
Instead, if using sieve of the sieve bottom under current mode with sieving wall vertical connected fashion, delay can be produced in perpendicular joints
The problem of accumulation.On the other hand, sieve wall height takes 3~5cm, and main function is the splashing for blocking liquid stream, and with the work of drainage
With.
But due to being washed by water in needing toward sieve in screening process, two-stage bolter also has difference.By
Belong to middle separation process, it is possible to which water injection system need not be set on sieve in first order separation；And the second level separates
Sieve belongs to ultimate separation, so needing to be washed away, in order to avoid leaving nematode, water spray system is must be provided with the bolter of the second level
System.It is of course also possible to set water injection system in first order separation, now the structure of two-stage bolter will be the same.
Because bolter needs water spray to wash away, the bolter for being provided with water injection system, bolter and water injection system
Structure design includes following several schemes：
Scheme one：
1st, in advance by the whole materials for sieving wall of enclosing more than sieve wall height, along height along the material edge sieve wall for being then more than this part
Then (the high 3~5cm of sieve wall) is first horizontal at degree rolls over straight down again to inboard folding is sieved, a circle shelves water plate 43 is formed in sieve,
As shown in figure 5, the high about 1-1.5cm of water fender；
2nd, in sieve wall top (upper along down about 1cm at, herein height of the height less than water fender), shelves water plate 43 pair
The sieve wall region progress answered is pre-perforated, and top sparge pipe 5 is bundled for wire rope handling；
Or carried out on shelves water plate 43 it is pre-perforated, for wire rope handling binding top sparge pipe 5 also may be used；
3rd, at inclined sieve to low spot, open up one and draw liquid mouth 44, draw and connect collecting pipe below liquid mouth, draw liquid mouth 44
Relative with 43 positions of shelves water plate, water fender is located at the high-end place of sieve, draws the lower end that liquid mouth is located at sieve；
4th, water injection system includes top sparge pipe 5, and top sparge pipe 5 can be both tied up on sieve wall into whole circle, can also be into
Whole circle is tied up on water fender (corresponding 2nd article two schemes)；Further say, top sparge pipe 5 is tied up when sieving on wall, also may be used
To be in wall or outside wall；It is preferable that way is generally to be bundled in inner side, because carrying apopore on sparge pipe, it is necessary to sieve
Interior water spray, if pipe is arranged on the outside of sieve wall, needs to open up inlet opening on sieve wall, can so increase the work for making sieve
Measure, if be bundled on the inside of sieve wall, the work for throwing water hole into can be saved, water pipe water outlet is directly entered in sieve.
Scheme two：
1st, as shown in fig. 6, half-turn sieve wall is made into a grade water plate 43 in advance, the high about 1-1.5cm of water fender, method is the same as scheme one
The 1st article；Water fender is positioned at the high-end position for tilting sieve；
2nd, in sieve wall top (upper along down about 1cm at, herein height of the height less than water fender), shelves water plate 43 pair
The sieve wall region progress answered is pre-perforated, and top sparge pipe 5 is bundled for wire rope handling；
Or carried out on shelves water plate 43 it is pre-perforated, for wire rope handling binding top sparge pipe 5 also may be used；
3rd, at inclined sieve to low spot, open up one and draw liquid mouth 44, draw and connect collecting pipe below liquid mouth, draw liquid mouth 44
Relative with 43 positions of shelves water plate, water fender is located at the high-end place of sieve, draws the lower end that liquid mouth is located at sieve.
4th, water injection system includes top sparge pipe 5, and top sparge pipe 5 can both be tied up in the sieve relative with water fender into half-turn
On wall, it can also be tied up into half-turn on the water fender；Further say, top sparge pipe 5 tie up sieve wall on when or
In wall or outside wall；It is preferable that way is generally to be bundled in inner side, reason is the same as scheme one；
5th, it is pre-perforated along the whole circle of sieve wall at sieve wall bottom (sieve bottom is up about at 1cm), bundle bottom sparge pipe for wire rope handling
6；
6th, water injection system also includes bottom sparge pipe 6, and bottom sparge pipe 6 is tied up into whole circle and sieving wall bottom, further, under
Portion's sparge pipe 6 can also be tied up on the inside of sieve wall, can also be tied up on the outside of sieve wall；It is preferable that way is in being generally bundled in
Side, because carrying apopore on sparge pipe, it is necessary to be sprayed water into sieve, if pipe is arranged on the outside of sieve wall, need in sieve wall
On open up water inlet, can so increase make sieve workload, if be bundled in sieve wall on the inside of, can save and throw water hole into
Work, water pipe water outlet is directly entered in sieve.
In above scheme, top sparge pipe 5, bottom sparge pipe 6, structure is as shown in fig. 7, it is about 5mm that water pipe, which is internal diameter,
Silicone tube, positioned at the top for tilting sieve, diameter is about 3mm for the water inlet 51,61 of pipe, for connect plus wetting system such as
Water pistol, pipettor or wash bottle etc..It is spaced apart on sparge pipe and is provided with several apopores 52,62, diameter about 1mm, towards sieve
Inner part is sprayed water.Add wetting system to carry out water filling to water pipe by water inlet 51,61, ejected further through apopore 52,62.
Top sparge pipe 5 is bundled on distance sieve wall along about 1cm at, the highly about 1.5cm of water fender 43, top sparge pipe 5
It can not only be bundled on sieve wall, it is preferred that top sparge pipe 5 can also be bundled on water fender, be bundled on water fender, court
To sieve wall water spray, such water can directly flow down along sieve wall.Water fender 43 can play a part of adjusting water (flow) direction, when top is sprayed
The water that water pipe sprays can be blocked when encountering water fender by water fender, and the water for spraying water pipe causes baffling, be gone downstream along sieve wall.
No matter which kind of binding mode water pipe takes above, its purpose is all current is flowed down along sieve wall, so as to wash away from top to bottom
Wall is sieved, the nematode being bonded on sieve wall is washed away.
Bottom sparge pipe 6 is bundled at the 1cm at sieve wall distance sieve bottom, around a circle.It is arranged at intervals on the sparge pipe of bottom
There are some apopores 62, diameter about 1mm, towards the position in sieve center；In bottom, the sieve wall peak position of sparge pipe opens up one
Water inlet 61, diameter are about 3mm, add wetting system such as water pistol, pipettor or wash bottle etc. for connecting；Add wetting system by entering greatly
The mouth of a river 62 carries out water filling to top water injection pipe, is ejected further through apopore 62.Preferably, bottom sparge pipe 6 leaves sieve bottom
Certain height, can so avoid bottom sparge pipe Shai Di corners because its stop and caused by remain in a standstill.
In such scheme, set diameter about 7mm's to draw liquid mouth 44 at the minimum point of sieve, be with draw that liquid mouth is connected
One collecting pipe 7, length about 5cm.Further, valve (when metal tube of the collecting pipe for hardness) can be set on collecting pipe 7, or
Clip (when collecting pipe is flexible pipe), collecting pipe 7 are mainly to shift the nematode suspension above bolter, and valve or clip can control
Nematode suspension enters collecting pipe.Connection collection vessel 8 below collecting pipe.
After I and II separates, the nematode suspension of reasonable particle is substantially achieved, is carried for nematode in nematode suspension
Take, mainly extracted using sucrose solution, refer to the separation described below and extracting method.
Package unit, including above-mentioned soil cup, bolter, water injection system, all supported by support and fixed.Especially exist
On support, bolter support is set, bolter support is the quoit or vinyon coil support of chamfer cylinder shape, such as
Shown in Fig. 1, sieve is supported, so as to form inclined bolter.
Two, the present invention also teaches the method using said apparatus separation and Extraction nematode, can be divided into following steps：
1st, macroscopic chad, fragment of brick and organic remains etc. are removed to carrying out simple process for examination soil first
Debris.
2nd, being put into for trying fresh soil in soil cup for 100g is weighed, 400ml distilled water or running water is added, starts
Agitator, the speed turned with 50-100 per minute, stir 5 minutes；According to soil texture situation, the proportion of sand to clay
For 1：2~1：5；
3rd, plug valve is opened above, opens collecting pipe below, is gone out the native liquid stream in soil cup, is passed through the following first order
The bolter of 40 mesh and the bolter of the mesh of the second level 400；
The native liquid on observation screen cloth is paid attention at any time, by controlling plug valve, is controlled the accumulating rate of native liquid at any time, is avoided soil
Hydrorrhea goes out the bolter of 400 mesh；
4th, it when native liquid is run up to above 400 mesh bolters, can be gently mixed with glass bar, while open and add wetting system,
Add water to upper and lower part sparge pipe, water is washed away sieve wall and sieve bottom, native liquid is flowed to liquid mouth direction is drawn；
5th, the residual of native liquid in soil cup is checked, is rinsed with a small amount of water, until being remained without native liquid, is then shut off plug valve.
When rinsing soil cup, water can constantly rinse 40 mesh bolters, untill observing through the native liquid clarification of 40 mesh bolters.
The purpose is to the nematode on bulky grain is flushed in 400 mesh bolters；
6th, constantly use plus wetting system is by water inlet, add water to top and the bottom sparge pipe, flushing is accumulated on 400 mesh bolters
The native liquid in face；
Observation passes through the native liquid of 400 mesh bolters, because the part of transmission contains all clay and fine silt,
In the case of continual rinsing, native liquid can also become to clarify, repeatedly with plus wetting system by water inlet to sparge pipe add water rinse sieve
Bottom, sieve wall is rinsed to top sparge pipe by water inlet, untill observing through the native liquid clarification of 400 mesh bolters；
7th, collection vessel is collected into nematode suspension, and by the precipitation of about 1 hour, centrifuge tube is transferred to after removing supernatant liquor
In；Collection vessel is rinsed well with water, suspension is centrifuged, 2000~3000 rev/min, then removes supernatant again
Liquid.
8th, 800g/L sucrose solutions are added in centrifuge tube, are centrifuged 2 minutes under 2000 revs/min of speed, by supernatant
The sieve noted into 500 mesh is sieved, and can obtain clean nematode with water flushing sieve.
It can be seen from above-described embodiment that the device taken of the present invention can realize soil nematodes be sufficiently separated and
Reduce the loss of nematode.
Claims (6)
1. a kind of device of soil nematodes separation and Extraction, including support, soil cup, agitator, bolter, water injection system, collection
Container, it is characterised in that：
The soil cup is the cup that a bottom has a liquid outlet at least, and a mozzle, water conservancy diversion are connected on the liquid outlet
One plug valve is set on pipe；
The agitator is placed in the soil cup；
The bolter includes first order bolter and second level bolter, and first order bolter is horizontally or diagonally to place, the
The second-order separation sieve is then slant setting, and in the second level, the lower end of bolter, which is provided with, draws liquid mouth, draws and collection is connected below liquid mouth
Pipe, collecting pipe lower section connection collection vessel；First order bolter is 40 mesh, and second level bolter is 400 mesh；
The first order bolter and the second level bolter include sieve bottom and sieve wall two parts, and sieve wall is originated from along sieve base
So linking forms upwards, and sieve bottom forms circular arc transition with sieving the place that wall is connected；
The water injection system includes sparge pipe, and the water outlet of the sparge pipe sieves wall from high-end to low side along the second level bolter
Place is washed away；
Second level bolter sieve cornice has a water fender, and the water fender is along on the inside of upper horizontally sieve by sieve wall
Folding, then rolls over and is formed straight down again；
The water fender forms whole a circle or half-turn second level bolter is inboard, when for half-turn when positioned at inclined described the
The high-end place of the second-order separation sieve；
The sparge pipe includes top sparge pipe, and when the water fender is a whole circle, the top sparge pipe is right into a whole circle
Tie up with answering in the region between the water fender and sieve wall, when the water fender is half-turn, the top sparge pipe is into partly
Circle, accordingly tie up in the region between the water fender and sieve wall, carry apopore on the top sparge pipe, water outlet along
Sieve on the inside of wall to dirty；
When the water fender and the top sparge pipe are half-turn, the sparge pipe also includes bottom sparge pipe, the bottom
Sparge pipe is tied up into whole circle and carries apopore, water side on the bottom of second level bolter sieve wall, the bottom sparge pipe
To towards on the inside of sieve；
The top sparge pipe is bundled on second level bolter sieve wall along down at 1cm herein, and be highly less than described
The height of water fender；The bottom sparge pipe is bundled in apart from second level bolter sieve bottom up at 1cm；
The top sparge pipe and bottom water spray bore are 5mm, dispersed on pipe to be provided with several water outlets
Hole.
2. the device of soil nematodes separation and Extraction according to claim 1, it is characterised in that：The soil cup is a bottom
Tapered cup, the liquid outlet are arranged at tapered bottom.
3. the device of soil nematodes separation and Extraction according to claim 1, it is characterised in that：The agitator includes rotation
Axle and stirring wheel, stirring wheel are made up of several blades that radially ring week is fixed on the rotary shaft, every blade and level
The angled inclination in face, plane body or helical form curved body are shaped as per blade.
4. the device of soil nematodes separation and Extraction according to claim 1, it is characterised in that：The top sparge pipe is tied up
On the sieve wall, or tie up on the water fender.
5. the device of soil nematodes separation and Extraction according to claim 1, it is characterised in that：The first order bolter and
Second level bolter all be slant setting in the case of, incline direction on the contrary, and the second level bolter angle of inclination be more than
The first order bolter.
6. the separating and extracting process that a kind of device according to one of Claims 1 to 5 soil nematodes separation and Extraction is taken, its
It is characterised by, comprises the following steps：
1) macroscopic debris is removed to carrying out simple process for trying fresh soil first；
2) being put into for trying fresh soil in soil cup for 100g is weighed, 400ml distilled water or running water is added, starts agitator,
With 50~100 revs/min of speed, stir 5 minutes；
3) plug valve on soil cup is opened, the collecting pipe is opened, goes out the native liquid stream in soil cup, pass through following first
Level bolter and second level bolter, flowed out from collecting pipe, into collection vessel；
4) when native liquid is run up to above the bolter of the second level, it is gently mixed with glass bar, while opens and add wetting system, to water spray
Guan Jiashui, water is washed away sieve wall and sieve bottom, native liquid is flowed to liquid mouth direction is drawn；
5) residual of native liquid in soil cup is checked, is rinsed with a small amount of water, until being remained in soil cup without native liquid, first order bolter
Untill native liquid clarification under sieve, plug valve is then shut off；
6) constantly with wetting system is added, by adding water to sparge pipe, flushing is accumulated in the native liquid above the bolter of the second level；Observation is saturating
The native liquid of second level bolter is crossed, untill through the clarification of the native liquid of bolter；
7) collection vessel is collected into nematode suspension, by precipitation, is transferred to after removing supernatant liquor in centrifuge tube, collection vessel
Rinsed well with water, suspension is centrifuged, 2000~3000 rev/min, then remove supernatant again；
8) in centrifuge tube add 800g/L sucrose solutions, centrifuged 2 minutes under 2000 revs/min of speed, by supernatant note into
The sieve of 500 mesh is sieved, and can obtain clean nematode with water flushing sieve.
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| CN105969654B (en) * | 2016-06-01 | 2018-08-03 | 延安大学 | A kind of fungal spore washing and sieving filter and fungal spore separation method |
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| CN113960285B (en) * | 2021-09-27 | 2023-11-17 | 中国热带农业科学院热带作物品种资源研究所 | Portable soil pathogen nematode integrated monitoring system |
| CN115901413B (en) * | 2022-12-23 | 2023-07-11 | 吉林省农业科学院 | Soil particle organic carbon test pretreatment device |
| CN116037555A (en) * | 2022-12-26 | 2023-05-02 | 生态环境部长江流域生态环境监督管理局生态环境监测与科学研究中心 | Benthonic animal panning device for fine sand substrate |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6019227A (en) * | 1994-11-16 | 2000-02-01 | May, Iii; Alexander Douglas | Extractor and separator apparatus |
| CN201308838Y (en) * | 2008-10-15 | 2009-09-16 | 云南农业大学 | Device for separating, collecting and counting soil nematodes |
| CN102628760A (en) * | 2012-03-31 | 2012-08-08 | 江苏徐淮地区淮阴农业科学研究所 | Cup-shaped multi-stage nematode separator |
| CN204034386U (en) * | 2014-09-03 | 2014-12-24 | 黑龙江省农业科学院大庆分院 | A kind of soil nematodes separator |
-
2015
- 2015-01-15 CN CN201510019727.9A patent/CN104568563B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6019227A (en) * | 1994-11-16 | 2000-02-01 | May, Iii; Alexander Douglas | Extractor and separator apparatus |
| CN201308838Y (en) * | 2008-10-15 | 2009-09-16 | 云南农业大学 | Device for separating, collecting and counting soil nematodes |
| CN102628760A (en) * | 2012-03-31 | 2012-08-08 | 江苏徐淮地区淮阴农业科学研究所 | Cup-shaped multi-stage nematode separator |
| CN204034386U (en) * | 2014-09-03 | 2014-12-24 | 黑龙江省农业科学院大庆分院 | A kind of soil nematodes separator |
Non-Patent Citations (3)
| Title |
|---|
| 不同种类杀虫剂对线虫的选择毒性研究;彭志国等;《垦殖与稻作》;20030831(第4期);第1.2.2节 * |
| 大豆胞囊线虫抑制性土壤及其抑制因子研究;宋洁;《中国优秀硕士学位论文全文数据库农业科技辑》;20130115(第01期);第11页第2.2.3节 * |
| 大连樱桃根际寄生线虫种类的调查与鉴定;江晓帆等;《安徽农业科学》;20140930;第42卷(第9期);第1.2节 * |
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