CN1019258B - Nematode storage and transport - Google Patents
Nematode storage and transportInfo
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- CN1019258B CN1019258B CN 85101908 CN85101908A CN1019258B CN 1019258 B CN1019258 B CN 1019258B CN 85101908 CN85101908 CN 85101908 CN 85101908 A CN85101908 A CN 85101908A CN 1019258 B CN1019258 B CN 1019258B
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- nematode
- missible oil
- antibacterial agent
- entomopathogenic
- nellie
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Abstract
The present invention relates to a method for improving the storage and the transport of nematodes. On one hand, the present invention provides a method for storing the entomopathogenic nematodes, the missible oil of the grubs of the infected entomopathogenic nematodes is mixed with an adsorbent, and the missible oil is stored under the condition that the growth of the microbe is suppressed; on the other hand, the present invention provides another method for storing the entomopathogenic nematodes, and the grabs of the infectedc entomopathogenic nematodes are stored under the basically anaerobic condition.
Description
On the one hand, the present invention has set forth the storage procedures of the entomopathogenic nematode that is specially adapted to prevent and treat harmful insect.On the other hand, the present invention has set forth the container that is suitable for storing and transporting nematode.
Can prevent and treat in harmful insect and the process exploring the alternative agricultural insecticide of environmental influence minimum, present notice is turning to biologic product, the entomopathogenic nematode class, comprising this too Nellie nematode section (Steinernematidae) and different Rhabditidae (Hetror-habditidae), provide a kind of like this possibility.They are natural pathogens of insect.
The entomopathogenic nematode of finding in soil is the third phase to infect larva.It can find and penetrate larva and the adult of insect.These nematodes have distinctive long altogether bacterium: have a liking for the different Rhabditidae nematode of bacillus/luminous heterorhabditis indica (Xenorhabdusnematophilus/lumines-cens).They help kill insects.In insect, in case the immune system of nematode interference insect and discharge the common long bacterium of rapid propagation will be destroyed the haemocoele (blood of insect and tissue) of insect.The haemocoele that nematode begins with bacterium and part digestion is a food, and ripe and breeding.This nematode and bacterium can cause that together insect is dead in 24-72 hour.
The notion of controlling insect with entomopathogenic nematode is a large amount of cultivations that are based upon nematode, and can be effectively used on the basis of the harmful insect of nematode sensitivity.Since a large amount of cultivation existing problems of nematode, and can not store entomopathogenic nematode effectively, and the development process that is used for the entomopathogenic nematode of biological control is hindered.In the past, the cultivation of nematode is limited to intracorporal method, as on greater wax moth (Galleria mellonella).But obtained remarkable progress with culture in vitro recently.Because it is realized artificial a large amount of nematode of cultivating, real more as the possibility of commercial biological insecticides with entomopathogenic nematode.
Before the practical alternatives of nematode as chemical insecticide, its storage and transportation have become key factor.
As everyone knows, the third phase nematode under extremely harsh condition (as dry and freezing), can be in soil the fine survival long duration.And when condition becomes favourable, can also recover, and when insect occurs, infect them.
Yet on the other hand, nematode is easy to damage for rapid drying with being under ultraviolet irradiation and the high temperature.
The third phase is infected the material existence of nematode larval (non-nursing phase) by storing in the body.
Probably, the growing period before desirable infection period, certain nematode survival ability of planting is relevant with the ability that its accumulates specific energy reserve (as glycogen or lipid), also with make them relevant with the ability of given pace activity.This speed is to satisfy nematode larval and do not grow the needs of (survival).In the third phase of infecting, in environment, be helpless to make the too fast movable of targeted insect host infection can continue the waste energy consumption, shorten survival period and infection chance subsequently.Therefore, the time-to-live partly can be by the decision of available energy conservation, but also may also comprise several other factors, as the generation and the secretion of the deactivation and the poisonous metabolic by product of enzyme.
Sell and be used for the nematode of the potency test of some insect at present, be stored under the low temperature (1-10 ℃).Under such condition, reduced the katabolism speed in the nematode.
Not being with continuous or method intermittently, is exactly to give airtight container oxygen supply by the nematode of limited quantity.Many workers emphasize that when storing nematode, the importance of oxidation and big surface area/volume ratio are to the importance of oxygen exchange.（Datky et al，1964 and Hara et al，1981）。Owing to use filter paper, test paper, velveteen and sponge etc., improved the contact area of nematode suspension and air.For example, on 100 gram cleaning dry sponge, 500,000,000 hair lines this too Nellie nematode (Steinernema feltiae) and march fly this too Nellie nematode (Sbibionis) can be at 1-2 ℃, store in the air bag of continuous charge to reach 4 months (Bedding, 1984).
Above-mentioned worker is providing the article of nematode that the preservation of nematode has been discussed for laboratory and field trial.
Usually because following shortcoming, described, be used for field trial and breadboard nematode storage procedures all is not suitable for wide range of commercial mould rule dispensing.
For example: (ⅰ) need be freezing
(ⅱ) need oxygen (it provides by the space of inflation or expansion inflation)
Even if (ⅲ) use measure above-mentioned, nematode density still limited (being up to 150000/milliliter).
Even if (ⅳ) use a year thalline, still be difficult to the extracting nematode.
One of purpose of the present invention is to provide the nematode that is more suitable in commerce is used to store and transportation resources.
The content of one aspect of the invention be with infectious nematode in anoxic be lower than under 40 ℃ the condition, this surprising discovery for a long time of still surviving is for the basis.This content on the one hand according to the present invention provides the method for storing entomopathogenic nematode, and wherein the missible oil of infectious entomopathogenic nematode larvae is stored under the condition of anaerobism basically.
" condition of anaerobism basically " this term meaning is that under these conditions, nematode can not get free oxygen.Be noted that in storage atmosphere the quantity of the oxygen of existence may change.The anaerobism effect may mean for nematode not of the same race different O
2Content.This content on the one hand of the present invention is with the preceding topic of being found to be of present inventor.Be that infectious larva does not need oxidation and ventilation or high surface/volume at lay up period.Therefore, initial storage requirement may be quite aerobic.In case but O
2Content is inhaled the effect of exhaling by nematode and is consumed, and it will become anaerobic condition.
Anaerobic condition can in all sorts of ways and reach.For example, propose as top, nematode itself can consume the oxygen in the storage atmosphere.On the other hand, nematode can be adorned envelope under vacuum, nitrogen, carbonic acid gas or other inert gas conditions.
Oxygen content is preferably less than the 0.4%(body), particularly less than the 0.25%(body).But just as set forth above like that, for nematode not of the same race, anaerobiosis may be higher than above-mentioned oxygen concentration.
Above-mentioned condition is relevant with the nematode population of commercial use, and promptly every gram missible oil contains 0.2 * 106 nematode at least.According to the technology (nematode concentration is quite low) of former description, this concentration is very surprising.
Nematode is preferably in this growth of microorganism and is subjected to store under the rejection condition, particularly under the condition that antibacterial agent exists.
This term of antibacterial agent used herein refers to and can destroy the material that bacterium and conk maybe can suppress bacterium and conk.It comprises bacteriostat, bactericide, fungistat and fungicide.
Antibacterial agent is formaldehyde or aqueous sodium hypochlorite solution preferably, and it is heavy that concentration is preferably 0.1-1.0%().There are many alternative antibacterial agents to use, as long as they are nontoxic to nematode in the quantitative range that uses.
According to another program of the present invention, before nematode is stored in the container, earlier with the antibacterial agent washing that can make the nematode surface sterilizing.
In the alternative content on the other hand of the present invention, provide a kind of so characteristic method.In the method, nematode stores in the solution that high penetration is arranged nontoxic to nematode and transports.In the optimal form of this scheme, it is that 30-70%(is heavy that nematode is suspended in concentration) monose or two sugar juices in.For example, sucrose or glucose.Because this principle without limits, can think that the solution of high penetration has reduced the undesirable growth rate of nematode.
In the content of another aspect of the present invention, nematode stores in the presence of adsorbent (as active carbon) or synthetic resin (as Amberlite).Adsorbent preferably is powdery, puts into the powder or the tablet of gel.In this situation of the present invention, nematode missible oil or suspension mix with an amount of adsorbent (as active carbon), and store under the condition that can suppress the nematode growth.In this case, nematode can store under anaerobic condition or aerobic condition or the vacuum basically.Nematode is preferably in the container that is in anaerobic condition basically and stores.Though anaerobic condition helps to reduce the possibility of the undesirable growth of nematode, it is found that the existence of oxygen does not have great influence.In nematode-adsorbent mixtures, it is heavy that adsorbent is preferably 10-70%().Draw thus, the 90-30%(that nematode missible oil is preferably mixture is heavy).In nematode missible oil, the concentration of nematode is preferably (0.2-2) * 10
6Individual nematode/gram missible oil.
Adsorbent is the lip-deep material that can be adsorbed onto various chemical substances it.So just can distinguish the material of adsorbent or other relative inertness (as wood chip, sponge, sand, filter paper and charcoal non activated).
Above-mentioned these methods have set forth that nematode stores and the new method of transportation, yet the present invention's content on the other hand not only according to the present invention but also according to the method for known storage nematode, has been set forth and stored and the container of transportation nematode.
Past, transportation is ordinary affair the nematode of carrying (as polyurethane sponge bits, filter paper, test paper, glass fiber or wood chip etc.) on the thalline matrix, so that improve total surface area/volume ratio, improve the transfer of the nematode ambient air that is contained in airtight container or tapping vessel.This content on the one hand according to the present invention, nematode is stored on powdered absorbent or the granule adsorbent.Under the situation of activated carbon powder, nematode can mix with active carbon, but under other many situations, promptly adsorbent is graininess or transports under the situation of carrying the nematode on the thalline matrix, before nematode was used for desirable place, it was necessary reclaiming nematode from carry thalline matrix.Reclaiming nematode may be consuming time many, and labour intensity is big, and usually causes that owing to the yield that removes denematize from carry thalline matrix is low nematode loses in a large number.In addition, some that exist in the nematode suspension that has separated are carried thalline matrix fractionlet and may be brought problem to post processing.For example, stop up used equipment.In order to address this problem, provide the container of storing and transport nematode here.This container is the airtight container that has two empty chambers and the equipment that is suitable for being retained in the above-mentioned first empty chamber to year thalline matrix and can allow nematode permeate to constitute.First sky chamber wiring worm is carried thalline matrix, and second empty chamber helps to reclaim nematode from above-mentioned matrix.Carrying equipment that thalline matrix is retained in the above-mentioned first empty chamber preferably by nematode sieve that can penetrate, that suitable sieve mesh is arranged, or the permeable sponge filter of nematode, or the sack of all or part of permeable nematode constitutes.
Though have manyly may belong to other form of the present invention, only describe several optimal cases now with reference to the accompanying drawings.Wherein:
Fig. 1 illustrates first optimal case of storing and transporting nematode.
Fig. 2 illustrates second optimal case storing and transport nematode.
Fig. 3 is the container profile of the 3rd optimal case of the present invention.
Fig. 4 is the container profile of the 4th optimal case of the present invention.
Fig. 5 is the container profile of the 5th optimal case of the present invention.
Fig. 6 is the container profile of the 6th optimal case of the present invention.
About Fig. 1.It has described closed container 2(such as plastic containers), it is equipped with the nematode missible oil 3 that mixes with adsorbent (as active carbon).The quantity of active carbon can change in very wide scope.But the quantity of active carbon is heavy at 30-50%() scope in, the result who is satisfied with.
About Fig. 2.Nematode missible oil 5 is contained in the semipermeable membrane air bag 4, for example dialysis tube.Air bag 4 is contained in the container 6, and this container is equipped with the active carbon suspension of active carbon or water or buffer solution.Antibacterial agent preferably is mixed in nematode missible oil or is mixed in container 6, capsule 4 liquid on every side.The scheme that Fig. 2 describes is particularly suitable for the situation that active carbon separates with the maintenance of nematode missible oil.The aperture of permeable membrane is preferably less than 10 microns.Therefore, on the other hand, provide the nematode bunkerage here, it is made of the semipermeable membrane air bag that can be contained in the sealing in the sealable container.Very thin powdered absorbent is housed between semipermeable membrane air bag and chamber wall or is dispersed in adsorbent in the aqueous solution.Adsorbent is active carbon preferably.The prescription that should be known in active carbon can make nematode load with high density, this Nellie nematode too of 1,000,000 hair lines of promptly every gram.To the size of packing without limits, can 1 gram following to more than 1 kilogram or 1 kilogram, this depends on the area of application and the consumption of unit are.
Should be known in that each above-mentioned scheme of the present invention all can make nematode depress storage in various degree anaerobism and oxygen.Nematode can store in a vacuum, or stores in nitrogen, nitric oxide, carbonic acid gas or air.And storage requirement does not require low temperature.Can see, from about room temperature to 37 ℃, the nematode ability of can surviving for a long time.
Turn over and see Fig. 3.Container 10 is made of lid 12 airtight empty capsids 11.Bag 13 is suspended on and covers below 12, and it is equipped with nematode and carries thalline matrix 14.The part 15 of bag 13, permeable past of nematode, and the not porous of matrix that carries disease germs.
Lid 12 is equipped with the mouth of pipe 16, and water can enter bag 13 by the mouth of pipe 16, so as to carry thalline matrix 14 the extracting nematode.
In the use, pump into water by the mouth of pipe 16, nematode washes out from carry thalline matrix 14 by permeable part 15.Empty capsid 11 can be used to wiring worm cleaning solution.As an alternative way, lid 12 and bag 13 can take out from container, and the nematode that washes out from bag 13 is packed into another container that is fit to.
Show that now in another selectable scheme, in order to be easy to import water by covering 12, the mouth of pipe 16 can replace with the part (Weakened portion) or the sheet segment (pull tab portion) of attenuate.
Turn over and see Fig. 4.Container 20 is made of lid 22 airtight empty capsids 21.It is divided into two chambers by means of the permeable net 25 of nematode.First Room and second Room are respectively 23 and 24.
In the use, the nematode of support nematode culture is carried thalline matrix 26 and is sent in first Room 23 of airtight container 20.When needs extracting nematode, take off and cover 22, wash nematode with water and carry thalline matrix, the nematode of suspension enters second Room 24 by net 25.The mouth of pipe 27 is contained in Lower Half second Room 24 of housing 21, so that can emit the suspension nematode from container 20.On the other hand, the mouth of pipe can replace with the part or the sheet segment of attenuate.In order to discharge the nematode that is suspended in the washings, can pierce through container.
Turn over and see Fig. 5, container 30 is made of lid 32 airtight empty capsids 31.The permeable bag 33 of nematode is placed in the empty capsid 31.It can make year thalline matrix 34 be retained in the bag 33, and makes the nematode of water and suspension pass through bag 33.
In use, can only take out bag 33 from container, wash out nematode.Or take off and cover 32, water washs by the bag 33 that carries thalline matrix 34 is housed.The part 35 of attenuate preferably is contained in the bottom of empty capsid 31, but so that provides the outlet of emptying for the nematode of water and suspension.
Turn over and see Fig. 6.Container 40 is for covering 42 airtight empty capsids 41.In the bottom of empty capsid 41 filler 43 being housed is the coarse grain foam material, as polyurethane sponge.Carry thalline matrix 44 and place the top of the porous spongy material 43 of coarse grain.In the use, take off and cover 42 or adorn a mouth of pipe covering, water washs through carrying thalline matrix 44, and extracting goes out nematode from carry thalline matrix.The nematode of water and suspension arrives aperture 45 through coarse grain foam material 43.Aperture 45 can be sheet segment, but then, can be made up of the material that is suitable for piercing through in the part of the attenuate of container bottom.
Previously described several schemes of the present invention and can in scope of the present invention, accomplish for the obvious improvement of these technology.For example, by suitable ram device, can make from first Room taking-up nematode of container and become more convenient.The method that another is desirable, container can have a common flexible outlet hose, discharge the second indoor water by the flexible pipe available pressure.
Above-mentioned these methods can be used for various nematodes, particularly pure nematode or this be the mixture of Nellie nematode section and different Rhabditidae bacterial classification too, comprising this Nellie nematode, Ge Shi Si this Nellie nematode (S.feltiae too of Nellie nematode (S.glaseri), hair line too too of march fly; Be the new parasitic nematode of steinernema [Neoaplectama carpocapsae] in the past, had a liking for bacillus heterorhabditis indica (Heteror habditis bacteriophora) and noctuid heterorhabditis indica (H.heliothidis).
Container is used at the scene that is useful for the above-mentioned transportation resources of the present invention, can be made and any suitable size arranged by any suitable material.For example, the mold of vacuum-packed plastics sachet, syringe-like, the bottle that is filled with air or indifferent gas, jar, tin can or dialysis tube.Nematode can directly be taken out the method for use or water or other solution dilution from container and be taken out use, and this depends on certain products prescription and application.
Be noted that the performance of estimate storing prescription is based on the infection ability of the nematode that stores and motility (survival ability).
Be used in the conventional bioassay of the nematode that the sheep bluebottle lucilia cuprina larva on the filter paper stores.Hatching was dissected larva and is determined whether they infect after 5 days in 23 ℃ of dark.The result who obtains compares with the result who is obtained by fresh nematode again.
In order briefly to describe the feature of invention, some specific examples are described now, only as an explanation.
Example 1
Ge Shi Si cultivates in the solitary parasitism culture of Nellie nematode strain NC513 on artificial dielectric very much, and this artificial dielectric is made up of chicken discarded object tissue homogenate and polyurethane sponge.From the third phase infection larva that artificial dielectric is gathered in the crops, and wash with water.The nematode of cleaning mixes with 0.1% formalin, and is stored in the air-tight bottle under 37 ℃.Experimental result provides in table 1.With water in contrast, the result proves and adds antibacterial agent (formaldehyde), increased the nematode bin stability.
In another identical experiment, storage is after 54 days down at 30 ℃, and the Ge Shi Si too survival ability of Nellie nematode is 90%.
Table 1 is stored too Nellie nematode existence of back antibacterial agent Dui Geshi Si down at 37 ℃
The influence of ability
The prescription survival ability
Ge Shi Si too the Nellie nematode in water after 13 days after 19 days after 30 days
- 0 -
Ge Shi Si too the Nellie nematode 0.1%
In the formalin 90 80 80
Example 2
As example 1 general introduction, preparation Ge Shi Si is Nellie nematode strain NC513 too.The nematode of having washed is suspended in M9 and dashes in the slow solution, and storing 30 days in the bottle in sealing under 22,28,33 and 37 ℃, in order to contrast, the nematode that is suspended in the water stored 19 days.
Provided the result of these tests at table 3.They show that physiology dashes the bin stability that the adding of delaying agent M9 has increased nematode.
Table 3 Ge Shi Si too Nellie nematode stores under different temperatures and (cushioned at M9 in 30 days
In the agent) after survival ability
Survival ability % after 19 days (water) and 30 days (M9)
Fill a prescription 22 ℃ 28 ℃ 33 ℃ 37 ℃
Ge Shi Si too the Nellie nematode in water 0000
Ge Shi Si too the Nellie nematode at the M9 buffer
*90 60 20-
*Consisting of of M9 buffer agent solution: in 1 liter distilled water, contain:
Na
2HPO
46 grams
KH
2PO
43 grams
Nacl 5 grams
MgSO
47H
2O 0.25 gram
Example 3
As example 1 general introduction, this Nellie nematode strain too of preparation Agriotis hair line.Nematode is suspended in water or 1.0% formalin, filters, and covers a little with activated carbon powder.Nematode is 22 ℃ of storages then.Table 2 is listed the result of the test of storing after 33 days.
These results have proved that adding adsorbent (as active carbon) has improved the bin stability of nematode greatly, and have proved by adding antibacterial agent (formaldehyde) and further improved stability.
Table 2 adsorbent and antibacterial agent to the hair line this too the Nellie nematode stored 33 days at 22 ℃
The influence of back survival ability
Prescription survival ability (%)
The hair line this too the Nellie nematode in water 0
The hair line this too the Nellie nematode in water+active carbon 80
This Nellie nematode 90-95 in 1% formalin+active carbon too of hair line
Example 4
As example 1 general introduction, this Nellie nematode strain too of preparation Agriotis hair line.The nematode of having washed is suspended in the high penetration solution (30% sucrose solution), and 22 ℃ of storages.Store that the survival ability of nematode is 85% after 33 days.The comparative trial of water shows that nematode has not had survival ability.
Example 5
As example 1 general introduction, this Nellie nematode strain too of preparation Otio hair line.The nematode of having washed (in every milliliter 0.1% formalin 4000) and activated carbon powder (in 0.1% formalin, concentration is 20% (weight)) were by mixing in 1: 1.
10 ml mixtures filter through giving the filter microcellular filler.The filler that has nematode and activated carbon powder that drains is fully put into the plastics sachet, and airtight.
After at room temperature storing 140 days, the survival ability of nematode is 92%.Store the postoperative infection ability decline is arranged slightly.
Example 6
As example 1 general introduction, this Nellie nematode strain too of preparation Mexican hair line.Contain 65 * 10
6The missible oil of individual nematode of having washed (has 1.5 * 10 in every milliliter 0.1% formalin
6Individual nematode) mixes by 1: 1 with the activated carbon powder of doing (diameter is less than 400 microns), be sealed in then in the plastics sachet.
Stored 30 days down and stored at least 180 days down at 4 ℃ at 24 ℃, infection ability is still intact.
Mixture in the sachet is suspended in the water, and is sprayed onto on the artichoke by high-pressure nozzle (2000 kPas).And draw, active carbon does not have injurious effects in application process.The temperature of storing nematode is satisfied to onion plume moth (artichoke plume moth, platyptilia cazrduidactyla).
Mixture in anther sac (90 gram) is enough to cover 500-5000 rice
2Soil (unit are consumption 10
4-10
5Individual nematode/rice
2).
List of references
Dutky, SR, Thompson, JB, and Cantwell, GE(1964) the prolific technology of DD-136 nematode.J·Insect Pathol，6417-422;
Hara, AHet al.(1981) solitary parasitism of the new parasitic nematode of weiser steinernema (Neoaptectana carpocapsae weiser) entomopathogenic nematode on dog food/agar medium cultivated in a large number, Advances in Agricul-tural Technology, Wester Series, № 16 June.
Bedding RA(1984) the new parasitic nematode of insect parasitic nematode belongs to the large-scale production of the different little bar layer of and, stores and transportation Ann App Biol 104 117-120.
Claims (18)
1, a kind of nematode preparation of compositions method that is suitable for prolonging period of storage, this method comprises:
The missible oil of the infectious entomopathogenic nematode larvae of 30-90% (weight) is mixed with the adsorbent of 10-70% (weight) and the antibacterial agent of antimicrobial effective amount.
2, a kind of nematode preparation of compositions method that is suitable for prolonging period of storage, this method comprises:
With of the antibacterial agent washing of infectious entomopathogenic nematode larvae with antimicrobial effective amount;
Prepare nematode missible oil by washed tool appeal entomopathogenic nematode larvae;
30-90%(is heavy) the missible oil and 10-70%(weight of tool appeal entomopathogenic nematode larvae) adsorbent mixes.
3, according to the method for claim 1 or 2, wherein said adsorbent is selected from active carbon and synthetic resin.
4, according to the method for claim 1 or 2, wherein adsorbent is an active carbon, and its concentration is that 10～60%(is heavy), nematode missible oil concentration is that 40～90%(is heavy).
5, according to the method for claim 1 or 2, wherein the concentration of nematode is (0.2～2) * 10
6Individual nematode/gram nematode missible oil.
6, according to the method for claim 1 or 2, wherein nematode is too mixture of any bacterial classification of Nellie nematode section nematode and different Rhabditidae nematode of pure nematode or this.
7, according to the method for claim 6, wherein said bacterial classification be march fly this too Nellie nematode, Ge Shi Si too the Nellie nematode, the hair line this too the Nellie nematode, have a liking for bacillus heterorhabditis indica and noctuid heterorhabditis indica.
8, according to the method for claim 1 or 2, wherein said antibacterial agent is selected from formaldehyde and liquor natrii hypochloritis.
9, according to the method for claim 1 or 2, wherein said antibacterial agent comprises the formaldehyde or the liquor natrii hypochloritis of hyperosmosis power.
10, according to the method for claim 1 or 2, wherein said its concentration of antibacterial agent is that 0.1～1.0%(is heavy) formalin.
11, according to the method for claim 1 or 2, this method also comprises described composition is stored in the container of sealing.
12, according to the method for claim 1 or 2, this method also comprises stores described composition under the condition of anaerobism.
13, according to the method for claim 12, wherein container seals.
14, according to the method for claim 2, comprising the antibacterial agent and the 30-90%(of antimicrobial effective amount is heavy) missible oil with appeal entomopathogenic nematode larvae and 10-70%(heavy) adsorbent mix.
15, a kind of method of storing entomopathogenic nematode, the missible oil of appeal entomopathogenic nematode larvae of wherein having is stored under the condition of anaerobism basically.
16, according to the method for claim 15, the nematode before wherein storing is washed with antibacterial agent.
17, according to the method for claim 15, wherein nematode stores under the condition that antibacterial agent exists.
18, according to claim 15,16 or 17 method, wherein missible oil is at least about containing 0.2 * 10
6Individual nematode/gram.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 85101908 CN1019258B (en) | 1985-04-01 | 1985-04-01 | Nematode storage and transport |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 85101908 CN1019258B (en) | 1985-04-01 | 1985-04-01 | Nematode storage and transport |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN85101908A CN85101908A (en) | 1987-01-10 |
| CN1019258B true CN1019258B (en) | 1992-12-02 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 85101908 Expired CN1019258B (en) | 1985-04-01 | 1985-04-01 | Nematode storage and transport |
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| CN101856015B (en) * | 2009-04-09 | 2013-01-16 | 中国科学院东北地理与农业生态研究所 | Method for refrigerated storage of heterorhabditis bacteriophora entomopathogenic nematodes in cold areas |
| CN106035238A (en) * | 2016-05-30 | 2016-10-26 | 中国科学院东北地理与农业生态研究所 | Nematode culture apparatus and simple breeding method for test entomopathogenic nematodes |
| CN107853290A (en) * | 2017-12-13 | 2018-03-30 | 朱郎平 | A kind of candidate stem cell freezes new method |
| WO2020047369A2 (en) * | 2018-08-30 | 2020-03-05 | The Curators Of The University Of Missouri | An efficient cryopreservation device preventing the direct contact between samples and extracelluar ice |
-
1985
- 1985-04-01 CN CN 85101908 patent/CN1019258B/en not_active Expired
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| CN85101908A (en) | 1987-01-10 |
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