EP0668718A4 - Verfahren zur lagerung von entomopathogenen nematoden. - Google Patents
Verfahren zur lagerung von entomopathogenen nematoden.Info
- Publication number
- EP0668718A4 EP0668718A4 EP93920592A EP93920592A EP0668718A4 EP 0668718 A4 EP0668718 A4 EP 0668718A4 EP 93920592 A EP93920592 A EP 93920592A EP 93920592 A EP93920592 A EP 93920592A EP 0668718 A4 EP0668718 A4 EP 0668718A4
- Authority
- EP
- European Patent Office
- Prior art keywords
- nematodes
- water
- mixture
- range
- container
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 241000244206 Nematoda Species 0.000 title claims abstract description 196
- 238000003860 storage Methods 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims description 49
- 230000000967 entomopathogenic effect Effects 0.000 title claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 80
- 239000002250 absorbent Substances 0.000 claims abstract description 54
- 230000002745 absorbent Effects 0.000 claims abstract description 46
- 230000000694 effects Effects 0.000 claims abstract description 42
- 239000000203 mixture Substances 0.000 claims abstract description 42
- 239000002245 particle Substances 0.000 claims abstract description 24
- 229920002401 polyacrylamide Polymers 0.000 claims abstract description 22
- 239000000463 material Substances 0.000 claims abstract description 20
- 230000001524 infective effect Effects 0.000 claims abstract description 12
- 241001523406 Heterorhabditis Species 0.000 claims abstract description 9
- 241001480238 Steinernema Species 0.000 claims abstract description 8
- 229940121375 antifungal agent Drugs 0.000 claims description 7
- 239000007863 gel particle Substances 0.000 claims description 7
- 239000003429 antifungal agent Substances 0.000 claims description 6
- 239000003242 anti bacterial agent Substances 0.000 claims description 5
- 229940088710 antibiotic agent Drugs 0.000 claims description 5
- 229920002472 Starch Polymers 0.000 claims description 4
- 241001465745 Steinernematidae Species 0.000 claims description 4
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 4
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 claims description 4
- 239000005864 Sulphur Substances 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000008107 starch Substances 0.000 claims description 4
- 235000019698 starch Nutrition 0.000 claims description 4
- 238000009423 ventilation Methods 0.000 claims description 4
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 claims description 3
- 241001465746 Heterorhabditidae Species 0.000 claims description 3
- 239000004021 humic acid Substances 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 2
- 239000000428 dust Substances 0.000 claims description 2
- 239000003077 lignite Substances 0.000 claims description 2
- 229920000609 methyl cellulose Polymers 0.000 claims description 2
- 239000001923 methylcellulose Substances 0.000 claims description 2
- 235000010981 methylcellulose Nutrition 0.000 claims description 2
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- 229920000058 polyacrylate Polymers 0.000 claims description 2
- 239000006071 cream Substances 0.000 abstract description 35
- 238000002156 mixing Methods 0.000 abstract description 10
- 230000004083 survival effect Effects 0.000 abstract description 3
- 230000007774 longterm Effects 0.000 abstract 1
- 239000000499 gel Substances 0.000 description 19
- 238000002474 experimental method Methods 0.000 description 15
- 241000894007 species Species 0.000 description 14
- 239000004927 clay Substances 0.000 description 11
- 239000002352 surface water Substances 0.000 description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- 241000238631 Hexapoda Species 0.000 description 8
- 241001480223 Steinernema carpocapsae Species 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 235000013305 food Nutrition 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 230000000366 juvenile effect Effects 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 241001523412 Heterorhabditis bacteriophora Species 0.000 description 4
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- 239000000047 product Substances 0.000 description 4
- 241001674044 Blattodea Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 3
- 229960000892 attapulgite Drugs 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- 239000004744 fabric Substances 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 229910052625 palygorskite Inorganic materials 0.000 description 3
- 229920000573 polyethylene Polymers 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 239000004721 Polyphenylene oxide Substances 0.000 description 2
- 229920005830 Polyurethane Foam Polymers 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000000274 adsorptive effect Effects 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 239000007900 aqueous suspension Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
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- 238000005187 foaming Methods 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
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- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229920000570 polyether Polymers 0.000 description 2
- 239000011496 polyurethane foam Substances 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
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- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 206010002660 Anoxia Diseases 0.000 description 1
- 241000976983 Anoxia Species 0.000 description 1
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- 229920000544 Gore-Tex Polymers 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 241000509371 Steinernema feltiae Species 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000607757 Xenorhabdus Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229940023476 agar Drugs 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 239000005030 aluminium foil Substances 0.000 description 1
- 230000007953 anoxia Effects 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 239000000688 bacterial toxin Substances 0.000 description 1
- 239000000679 carrageenan Substances 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 229940113118 carrageenan Drugs 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
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- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
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- 150000004676 glycans Chemical class 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000008240 homogeneous mixture Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 201000002266 mite infestation Diseases 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 229910052939 potassium sulfate Inorganic materials 0.000 description 1
- 239000001120 potassium sulphate Substances 0.000 description 1
- 235000011151 potassium sulphates Nutrition 0.000 description 1
- 239000011369 resultant mixture Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
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- 239000007921 spray Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000000196 tragacanth Substances 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Definitions
- This invention concerns the storage for transport or future use of entomopathogenic nematodes. More particularly it concerns the storage and transport of the third stage infective juveniles of nematodes belonging to the genera Steinernema (synonym Neoaplectana) and Heterorhabditis (synonym Chromonema) using highly absorptive substances, such as polyacrylamide gels, at specific water activities.
- Nematodes have also been mass produced in liquid culture as detailed by G W Pace et al (see WIPO Publication No 86/01074), and by M J Friedman, S L Langston and S Pollitt (see the specification of International patent application No PCT/US88/04124, which is WIPO Publication No WO 89/04602).
- G W Pace et al see WIPO Publication No 86/01074
- M J Friedman, S L Langston and S Pollitt see the specification of International patent application No PCT/US88/04124, which is WIPO Publication No WO 89/04602.
- Popiel et al store their nematodes at relative humidities in the range of from 50 to 94 per cent, or from 95 to 99 per cent, while allowing for adequate oxygen supply. These humidities are maintained (i) with a hydrogel or fibrous matrix that has been impregnated with a saturated solution of potassium sulphate (which maintains a relative humidity of 97 per cent), (ii) using a saturated solution of potassium nitrate (which has a relative humidity of 94 per cent), or (iii) using solutions of sulphuric acid.
- the preferred technique involves storing the desiccated nematodes in a container in which there is also a saturated salt solution in a package made from a hydrophobic, vapour permeable membrane, such as GORETEX (trade mark) material.
- the cryptobiotic nematodes are stored in an airtight and moisture tight container with sufficient air space to accommodate the needs of the stored infective juveniles.
- Popiel et al claim that, using this technique, nematodes of the species Steinernema carpocapsae, Steinernema feltiae (synonym bibionis) and Heterorhabditis bacteriophora (synonym heliothidis) survived for several months with undiminished infectivity.
- nematodes In order to remove surplus water, a thin layer of nematodes is subjected to vacuum filtration followed by air drying. This takes much space and time. In addition, it is possible that nematodes may be excessively dried, particularly under commercial production conditions.
- the thin layer of nematodes must be removed from the treatment site and placed in containers for storage, with special provisions for maintaining the relative humidity near, but below, 100 per cent.
- a hydrogel structure made from a polysaccharide (such as agar, carrageenan or tragacanth), or a porous matrix of sponge, polyurethane foam or polyether foam, contains the nematodes, which "nictate" or “stand up and wiggle" on a relatively dry surface provided on the surface of the nematode-containing medium.
- the relative humidity of the air surrounding the nematode-containing medium is maintained at a value of at least 95 per cent by enclosing, with the medium, a quantity of "a water-liberating gel, such as swelled polyacrylamide".
- R A Bedding showed how clay (particularly attapulgite clay) can be used to store entomopathogenic nematodes, either by forming a homogenous mixture of nematodes with the clay (which is not a highly absorbent material) or by making a sandwich consisting of a layer of nematode cream between two layers of clay.
- This method (which is currently used commercially) is believed to owe its success to three major factors, namely (i) the nematodes are constrained from moving (thus conserving food reserves), (ii) perhaps, the clay adsorbs potentially toxic nematode excretory products, and (iii) by exposing nematodes to humidities of less than 100 per cent, preferably near to a water activity of 0.97, they are induced into the cryptobiotic state.
- a range of entomopathogenic nematode species can be stored in this way using either chips of calcined attapulgite clay or coarsely milled, calcined attapulgite clay.
- More than half the weight of the stored nematode product is clay, which adds to transport costs when air freight is used.
- This objective is achieved by using highly absorptive (but not necessarily adsorptive) gels, or other highly absorptive substances, to absorb surface water from an aqueous cream of nematodes and, having done this, to accurately provide a water activity within a narrow range suitable for inducing cryptobiosis of the nematodes without adversely affecting their survival.
- Water activity is defined as the ratio RH/100, where RH would be the relative humidity of the surrounding atmosphere in a sealed system.
- highly absorptive substance will be understood by persons who work with absorptive materials and are familiar with their classification. Without limiting the general understanding of the term “highly absorptive”, the preferred highly water-absorptive substances of the present invention are substances which absorb at least 75 per cent of their own weight of water to have a water activity in the range of from 0.80 to 0.995
- anhydrous or nearly anhydrous absorbent particles are preferably mixed with a nematode cream in predetermined combinations so that both the absorption of the surface (free) water and the establishment of the desired water activity are achieved without requiring further adjustment of the resultant mixture (for example, the addition of extra water).
- Fungicides and/or antibiotics may be mixed with the nematode cream or with the absorbent prior to mixing the nematode cream and absorbent together.
- the benefits of using highly absorbent substances are believed to be five-fold. Firstly, they remove surface water rapidly so that the nematodes form a "foam" within a few minutes. Where particles of an absorbent (such as polyacrylamide gel) that swells in the presence of water are used, large interstitial spaces are formed between the swollen particles. These spaces provide room for nematode foaming and for the better diffusion of gases. Thus an aerated matrix is rapidly produced and the nematodes suffer the effects of anaerobic conditions for a short time only.
- an absorbent such as polyacrylamide gel
- the mixture not only induces cryptobiosis in the nematodes, but it also, thereafter, maintains the water activity at a value which is appropriate for the continual storage of the nematodes.
- the weight of absorbent used can be significantly less than that of the nematode cream.
- Nematodes stored in this manner using highly absorbent substances can be reactivated by dispersing the nematodes and absorbent in water. However, they are most effectively reactivated by adding water to the nematode/absorbent combination so that the water activity exceeds 0.995 but remains below 1.00; the nematode/absorbent combination being then left for two to four hours before finally dispersing the nematodes and absorbent in a surplus of water. Where particles of the absorbent swell in the presence of water (for example, when the absorbent is polyacrylamide gel), it is then a simple matter to sieve out those particles to leave a clean suspension of infective juvenile nematodes.
- a method of storing third stage infective juveniles (J3) of entomopathogenic nematodes comprises forming a mixture of an aqueous concentrate (cream) of clean J3 nematodes and particles of a highly water-absorbent (as defined above) material, the proportions of the aqueous concentrate and water-absorbent material being such that the mixture, after equilibrating, has a water activity in the range of from 0.92 to 0.995.
- the water activity of the mixture is from 0.92 to 0.995, and most preferably between 0.95 and 0.99.
- the final value of the water activity of the mixture may take from 24 to 72 hours to establish.
- the absorbent may comprise from about 15 per cent to 75 per cent of the resulting mixture.
- experiments should be conducted to determine the water activity resulting from various combinations of it with nematode cream.
- the water content of the nematode cream must first be standardised. This can be achieved by making the number of clean infective juvenile nematodes per gram constant for any particular species of nematode or by standardising the viscosity of the nematode cream used. The more viscous the nematode cream, the lower the quantity of absorbent that is required to achieve a particular water activity of the final absorbent/nematodes mixture.
- the exact water activity required to give optimal induction of cryptobiosis varies a little from species to species of entomopathogenic nematodes. After a period of induction (usually 2 to 3 days), the nematodes may be stored with the mixture having the attained water activity, or the water activity of the resulting combination may be lowered by adding further absorbent.
- PAGs particulate anhydrous polyacrylamide gels
- a PAG having an individual dry particle weight of from 0.005 gm to 0.02 gm is preferred because, after swelling, such particles provide adequate interstitial space for nematode foaming and gaseous diffusion.
- Figure 1 is a graph showing the water activity of various mixtures of polyacrylamide gel and water.
- Figure 2 is graphical presentation of mortality data for various absorbent/nematodes combinations of the nematode species Steinernema carpocapsae.
- Figure 3 is a graphical presentation of the average dry weight of nematodes (S. carpocapsae) in stored samples after different storage times.
- entomopathogenic J3 nematodes that have been reared in a liquid culture frequently store less well than those reared in a solid culture (this is particularly true of Heterorhabditis species).
- utilisation by the nematodes of their internal food reserves prior to storing should be reduced to a minimum.
- nematodes used in the series of trials conducted by the present inventors to confirm the efficacy of the present invention were reared and extracted using the methods described by R A Bedding, M S Stanfield and G W Crompton in the specification of International patent application No PCT/AU91/00136 (WIPO Publication No WO 91/15569).
- the nematodes may be reared on insects in vivo or in liquid culture, provided the nematodes are free from appreciable amounts of extraneous matter remaining from the culture medium and are relatively free from nematode stages other than J3 (preferably no adult nematodes are present and certainly no more than 2 per cent of the nematodes should be adults).
- the resulting cream of nematodes contained from 0.5 to 3.5 million J3 nematodes per gram, depending upon the species involved and the amount of inter-nematode water remaining.
- various antifungal and antibiotic agents were added to and mixed with the sedimented nematodes while they were still in the tanks. Hence, after removal of much of the surface water (and thus also of most of the antibiotic and/or antifungal agent), some of the antibiotic and/or antifungal agent remained to be absorbed by the absorbent in the storage process.
- the antifungal agents used by the present inventors were humic acid, brown coal dust (which contains up to about 50 per cent humic acid), powdered sulphur, sulphurous acid, and a mixture of sulphurous acid and powdered sulphur.
- the technique usually adopted for combining the nematodes and the absorbent material was as follows. Particles of the absorbent were weighed and added to the appropriate weight of nematodes. (The appropriate weights were determined by prior experimentation to ascertain which combinations fall within the required range of water activities). The absorbent and nematodes were then immediately stirred and mixed together so that the absorbent particles were evenly distributed in the mixture.
- the water activity, of the mixture of nematodes and highly water-absorbent particles has to be in the range of from 0.80 to 0.995.
- the water activity is preferably in the range of from 0.92 to 0.995, and most preferably is in the range of from 0.95 to 0.99.
- polyacrylamide gel is the preferred absorbent and Figure 1 shows the ratio of water to polyacrylamide gel to achieve a water activity for the mixture in the range of from 0.95 to 0.995.
- the required water activity of the mixture is not attained immediately.
- the absorbent particles quickly take up the free surface water of the nematode cream, and then absorb water that is released from within the nematodes.
- the absorbent material has a water activity lower than its final value, which is attained after 24 to 72 hours.
- the water activity of the gel particles is 0.92 for the first four hours after the mixing has taken place. The water activity then increases to 0.94 eight hours after mixing, but does not attain the (intended) value of 0.97 until 24 hours after the mixing of the nematode cream and the absorbent particles.
- the containers were stored at the extended storage temperature of the experiment immediately after receiving a sample of the mixture of nematodes and absorbent material. In other experiments, the containers were stored firstly at 15°C for three days and then at the extended storage temperature. In all experiments, the extended storage was effected in a manner such that the water activity of the absorbent/nematodes combination was maintained at a value in the range of from 0.80 to 0.99.
- the squeezed cream had a viscosity such that when it was placed in a 4 cm open ended tube held vertically, it did not drop out of the tube within 2 minutes. (The addition of even small quantities of water to some of this cream reduced its viscosity so that it readily dropped from the tube within a few seconds.
- the squeezed cream was used to determine what water activities were achieved with various combinations of sieved polyacrylamide gel crystals (the crystals having an average mass of 0.01 gm) and this species of nematode, and how these water activities affected nematode mortality if the combination is stored at 23°C.
- the nematode cream was divided into seven batches and combined with the PAG crystals in the following proportions by weight:
- Nematodes were produced and processed as in Example 1, but with 10% more fat added to the rearing medium. Batches of from 200 gm to 500 gm of an aqueous cream of nematodes were combined with particles of sieved PAG (average particle weight, 0.01 gm). Surface water in the nematode cream could not be standardised as readily as in Example 1 because Heterorhabditis bacteriophora cream loses water much more readily than Steinernema carpocapsae. Accordingly, the H. bacteriophora nematodes were standardised by making a thick cream and then diluting it with water, so that there were about 2.5 million infective third stage juvenile nematodes per gram. This diluted cream of nematodes was combined with polyacrylamide gel particles as in Example 1.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AUPL479192 | 1992-09-10 | ||
AUPL4791/92 | 1992-09-10 | ||
PCT/AU1993/000465 WO1994005150A1 (en) | 1992-09-10 | 1993-09-10 | Method for the storage of entomopathogenic nematodes |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0668718A1 EP0668718A1 (de) | 1995-08-30 |
EP0668718A4 true EP0668718A4 (de) | 1997-07-23 |
Family
ID=3776424
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP93920592A Withdrawn EP0668718A4 (de) | 1992-09-10 | 1993-09-10 | Verfahren zur lagerung von entomopathogenen nematoden. |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP0668718A4 (de) |
JP (1) | JPH08501536A (de) |
CN (1) | CN1088046A (de) |
CA (1) | CA2144172A1 (de) |
IL (1) | IL106980A (de) |
TW (1) | TW230186B (de) |
WO (1) | WO1994005150A1 (de) |
Cited By (1)
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CN106993607A (zh) * | 2017-04-28 | 2017-08-01 | 广东省生物资源应用研究所 | 腐殖土作为载体在贮存昆虫病原线虫中的应用 |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994019940A1 (en) * | 1993-03-04 | 1994-09-15 | Commonwealth Scientific And Industrial Research Organisation | Method for packaging entomopathogenic nematodes for storage and transport |
FR2744878B1 (fr) * | 1996-02-15 | 1998-04-17 | Lopuszanski Daniel | Recipient pour le conditionnement de vers marins pour la peche |
AUPP616798A0 (en) * | 1998-09-28 | 1998-10-22 | Commonwealth Scientific And Industrial Research Organisation | Method and apparatus for the storage of entomapathogenic nematodes |
US6399118B1 (en) * | 2001-06-29 | 2002-06-04 | Fish Biotech Ltd. | Process for storing enriched nematodes |
CN101149366B (zh) * | 2007-10-16 | 2015-03-04 | 李红玉 | 一种快速定量线虫死亡率的新方法 |
CN104222075B (zh) * | 2014-08-29 | 2015-12-09 | 上海大井生物工程有限公司 | 一种哈茨木霉菌菌剂载体的配方及其制备工艺 |
CN104222023B (zh) * | 2014-08-30 | 2017-07-25 | 北京安和亿泰生物工程技术有限公司 | 一种斯氏线虫大批量繁殖培育方法 |
US11013229B2 (en) | 2015-05-04 | 2021-05-25 | Green Advantage Technology Inc. | Methods and formulations for storing entomopathogenic nematodes |
CN105532583B (zh) * | 2015-12-29 | 2018-04-06 | 宁波市农业科学研究院 | 一种昆虫病原线虫的活体繁殖方法 |
CN113080186B (zh) * | 2021-03-18 | 2022-04-12 | 中国林业科学研究院林业新技术研究所 | 一种扩散型3龄松材线虫的低温保存方法 |
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JPS6422801A (en) * | 1987-07-16 | 1989-01-25 | Oji Paper Co | Preservation of nematode living in insect |
WO1992010170A1 (en) * | 1990-12-07 | 1992-06-25 | Temple University - Of The Commonwealth System Of Higher Education | Stabilized insect nematode compositions |
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Publication number | Priority date | Publication date | Assignee | Title |
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EP0256873B1 (de) * | 1986-08-18 | 1993-01-27 | Biosys | Kommerzielle Lagerung und Versendung entomogener Nematoden |
EP0359764B1 (de) * | 1987-05-05 | 1994-03-09 | Commonwealth Scientific And Industrial Research Organisation | Lagerung von entomopathogenen nematoden |
AU635921B2 (en) * | 1989-02-21 | 1993-04-08 | Biosys | Method and medium for packaging entomogenous nematodes |
US5172514A (en) * | 1990-11-19 | 1992-12-22 | Biosys Corporation | Insect trap |
AU665398B2 (en) * | 1992-02-28 | 1996-01-04 | Commonwealth Scientific And Industrial Research Organisation | Preparations containing entomopathogenic nematodes for biological control of the banana weevil |
-
1993
- 1993-09-10 IL IL10698093A patent/IL106980A/xx not_active IP Right Cessation
- 1993-09-10 CN CN93119099A patent/CN1088046A/zh active Pending
- 1993-09-10 EP EP93920592A patent/EP0668718A4/de not_active Withdrawn
- 1993-09-10 JP JP6506692A patent/JPH08501536A/ja active Pending
- 1993-09-10 CA CA002144172A patent/CA2144172A1/en not_active Abandoned
- 1993-09-10 WO PCT/AU1993/000465 patent/WO1994005150A1/en not_active Application Discontinuation
- 1993-09-27 TW TW082107955A patent/TW230186B/zh active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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JPS6422801A (en) * | 1987-07-16 | 1989-01-25 | Oji Paper Co | Preservation of nematode living in insect |
WO1992010170A1 (en) * | 1990-12-07 | 1992-06-25 | Temple University - Of The Commonwealth System Of Higher Education | Stabilized insect nematode compositions |
Non-Patent Citations (2)
Title |
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DATABASE WPI Section Ch Week 8910, Derwent World Patents Index; Class C03, AN 89-071638, XP002031700 * |
See also references of WO9405150A1 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106993607A (zh) * | 2017-04-28 | 2017-08-01 | 广东省生物资源应用研究所 | 腐殖土作为载体在贮存昆虫病原线虫中的应用 |
CN106993607B (zh) * | 2017-04-28 | 2020-10-20 | 广东省生物资源应用研究所 | 腐殖土作为载体在贮存昆虫病原线虫中的应用 |
Also Published As
Publication number | Publication date |
---|---|
IL106980A (en) | 1997-02-18 |
WO1994005150A1 (en) | 1994-03-17 |
TW230186B (de) | 1994-09-11 |
CN1088046A (zh) | 1994-06-22 |
JPH08501536A (ja) | 1996-02-20 |
CA2144172A1 (en) | 1994-03-17 |
EP0668718A1 (de) | 1995-08-30 |
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