BG1134U1 - Virus-alkaloid mixtures for insect control - Google Patents

Abstract

The utility model relates to insecticide mixtures, which consist of entomopathogenic viruses, total extract of alkaloid-bearing plants and oil, and are applicable for pest control in agriculture and forestry. The virus-alkaloid mixtures cause insects like apple fruitwarms (Laspeyresia pomonella L.), plant lice (Aphididae), etc. Perish; it combines the contact effect of the alkaloid and the viral infection effect to occur after a certain period of time (incubation period). The insecticide mixture consists of an entomopathogenic virus and total extract of Nicotiana tabacum, L. and/or Veratrum album L. in the following amounts in the working solution: entomopathogenic virus û 4 to 10 volume percent (selected from the group of Baculovirus); 0.5 to 0.65 weight percent of total tobacco extract or 0.8 to 1.0 weight percent of Veratrum album L., all ingredients being pre-dissolved in a mixture of soybean oil and Tween 80« and water.

Description

1134 IU (54) VIRUS-ALCALOID MIXTURES NOSE TREATMENT FIELD OF THE INVENTION The utility model relates to the application of insecticidal mixtures consisting of tetanopathogenic viruses, total extract of Nicotiana tabacum L and / or Veratrum album L. harboring plant pests in rural IGO. Viral alkaloid infections cause the perishing of insects, such as fruit worm, aphids, and the like, combining the contact activity of the alkaloids and the effect of viral infection for a prolonged period of time (incubation period). BACKGROUND OF THE INVENTION [0002] Viruses are known to consist of viruses only - polyvalent polyhedrosis virus (SU 1836005 A3, SU 2153258 et al.), Cabbage worms which consist solely of entomopathogenic nuclear polyhedrosis viruses. There are known viral preparations comprising mixtures of nuclear polyhedrosis viruses (SU 1637653) as well as baculovirus (nuclear polyhedrosis virus) in combination with fermenta chitinase (SU 2059368). An insect mixture consisting of a granulous virus which is precipitated with a basic protein (at pH = 8) after release of the virions with the ability to disperse grease is known (US 3716634). A viral insecticide comprising virons contacted with a polarized organic solution under hydration conditions in combination with a lipid which is readily dispersible is known (US 3639578). A viral preparation (SU 591119) - Switzerland - Sandoz AG, consisting of NPV - Heliothis zea and a matrix consisting of proteinaceous material or clay - spray-dried is known. There is known a viral composition of ultraviolet-protected insect viruses comprising a nuclear polyhedrosis virus, activated carbon and ethylcellulose (US 3541203). There are known viral insecticides consisting of entomopathogenic viruses in admixture with proteinaceous material or clay (Switzerland -619352). It is known to be a fungicide pest control agent for plants and animals which is a extract and tobacco powder (Nicotianatabacum L). The preparation is recommended for fighting saliva, ants and mushroom diseases (DE 4239222 A1). A mixture of vegetable oil (soybean, sunflower, rapeseed, cotton) and pyrethrum (containing alkaloidapyrethrine) is known, but piperenebutoxide (surfactant which does not have an insecticidal action) is used as an emulsifier for the vegetable oil (DE 4421417 A1). Insecticide mixtures for Adoxophyes orana (Lepidoptera, Tortricidae), consisting of NeemAzal T / S, a product comprising an extract of Azidirachta indica and entomopathogens, are known. Insecticide mixtures for aphids are used in apple gardens presenting a combination of Bacillus thuringiensis or granulosus virus to combat certain leukocytes. The benefits of the proposed blends are reflected in the following. When used to combat pests of an entomopathogenic virus or other pathogen, it is known that it first has to be swallowed and, after a certain incubation period, the insect infestation is multiplied, sepsis the disease and the insect perishes. Throughout this time the insect is eating and causing damage to the fruit or leaves growth that is not desirable. Literary data show that mortality of up to 80% of other living bugs of apple fruit worm (Jaques et al., 1987) was observed after 17 days of infection. When used to control only alkaloid-containing extracts, a contact action is observed in which the insects rotate rapidly. Under the influence of light, temperature and other abiotic factors of the environment, biologically active substances rapidly inactivate, which means they have a short-term effect. Insecticidal mixtures consisting of a pathogen and an alkaloid combine by their effect the factors that are useful for protecting growth, such as the rapid initial action of the alkaloid and the effect of the disease on the pathogen after a certain period of time. This way a very small percentage of the insects are left to eat, which subsequently perish from the pathogen. Technique of the utility model In order to expand the assortment of highly insecticidal preparations and to overcome the resistance of apple fruit juice against the granule virus on the apple flatworm (Eberle KE, JA Jehle, 2006) to use viral-alkaline mixtures (pH 6-8) consisting of an antropic virus and a total extract of Nicotiana tabacumL and / or Veratrum album L. to combat pests in agriculture and forestry, containing: An entomopathogenic virus - preferably a granulosa virus on apple fruit (9x109 granules / ml) in a quantity of 5 ml of a solution of 2-500 [mu] l and a total ethanol hunting tobacco extract (Nicotiana tabacum L) prepared according to the method described by Atanassov, Stoyanova 1997) but using 70% ethyl alcohol and an extract content of 0.1-6.5mg / ml or 2. Baculovirus Laspeyresia Granulosa virus prepared in 5 ml working solution in accordance with (1) and total extract of Cheat (Veratrumalbum L) prepared according to (1) and a content extract of 0,4-10 mg / ml or 3. Total tobacco extracts (Nicotianatabacum L) (Veratrum album L), and Baculovirus Laspeyresia gram-negative virus with a content of biologically active substances in quantities and prepared according to points 1 and 2. Examples of the utility model Preparation of the viral-alkaloid mixtures . Extracts of Nicotiana tabacum L. or Veratrum album L., prepared in the form of powdered or powdered pulp, are dissolved in a mixture of Twin 80: soybean oil (1: 1), after which the necessary quantity of distilled water with the granulosa virus . The individual constituents comprise for 100 ml of distilled water - N. tabacum extract - 0.0165-0.652%, V. album-0.005-0.8-1.0% and B. Laspeyresia - 0.49-2.120%. If it is necessary that the pH can be maintained with any of the known water-soluble phosphate compounds of potassium or sodium. The composition contains a large amount of viral granules. The viral insecticide used in the mixture relates to the genus Baculovirus - polyhedron gamma viruses. The virions are immersed in a bead matrix and contain from 1 to 1000, and more preferably in each polyhedron or 1 in the granulate. These viruses cause diseases of insects such as nuclear polyhedroses and granuloses, etc. Granulous viruses are preferably used in the mixtures, as well as nuclear polyolefins isolated from B. mamestra, B. Lymantrie et al. Experiment 1. Extracts of N. tabacum L. were prepared by the method described by Atanassov, Stoyanova (1997), but using 70% ethyl alcohol and tested their efficacy at the age of first-time caterpillars of apple fruit pomace (Laspeyresia pomonella L) using the method described by the same authors (Atanassov, Stoyanova, 1997a) (Table 1). Experiment 2. Veratrum album L. extract is prepared by the aforementioned method described in Example 1 and tested for efficacy with respect to the first age of apple fruit jelly according to Experiment 1 (Table 2). Experiment 3. Apple plum pest grain virus is prepared after purification in a charcoal gradient resulting in a vigorous suspension of 9 x 10 < 6 > / g / ml titre and assaying its effectiveness against first-age caterpillars of apple fruit worm (Table 3). After mathematical data processing using regression analysis and plotting regression lines (Figure 1) for each component, working tables 4 and 5 are developed. The tables are a tabular method, the effectiveness of the used plant extracts from tobacco or blackberries to combat first-age caterpillars of apple fruit worm applied alone or as mixtures of gram-negative viruses on the apple fruit worm. 3 1134 W The prepared tables allow theoretically to see the increase in mortality up to 16th of the infection with the virus. Example 1 Tobacco extract (Nicotianatabacum L.) 5 Preparation of a solution containing 10 mg / ml of extract is carried out by measuring 250 mg of tobacco extract prepared as a capsule and dissolving it with 500-1000 microlitres of Twin- 80: 500-1000 microl of soybean oil (1: 1) and add 10 ml of 20 ml of distilled water. A solution containing 10 mg / ml of extract is obtained from which we obtain the other concentrations - 5 mg / ml, and so on. The extract is tested by sprinkling the apple fruits, and after the solution is dried, we place the fruit in plastic buckets with a latch (200 ml) and infect them with two first-time caterpillars of apple-like worm. The experience is set in four repetitions with 15 apples in each. The results obtained are reported in Table 1 and Table 1A. Table 1 A Variants Amount of insects Concentration of insecticide Mortality of insects per pcs. % w / v% Tobacco extract 60 0.5 5 100 60 0.3 3 86 Super Sect 58 0.02 100 Control - distilled water 60 - 0 Example 2 Chelate extract (Veratrum 25album L.) The working solution is obtained we measure 200 mg of a tea extract prepared as a powdered or mash and dissolve it in 500-1000 microl Twin 80®: 500-1000 microliters of soybean oil 30 (1: 1), then add 20 ml of distilled water. A suspension is obtained with an extract content of 10 mg / ml. It is prepared by dissolution and solutions with a content of 5 mg / ml and the like. The test of the insecticidal action of extracts against caterpillars first apple fruit worm grows as example 1. The statistical processing of the data and the best results are shown in Nos. 2 and 2A. Table 2 A Variants Amount of insects Concentration of insecticide Insect mortality of § £% mg / ml% Chew extract 60 1.0 10 100 «6 60 0.8 8 97 59 0.5 5 90 60 0.3 3 79 Supersec 75 , 02 100 Control - distilled water 60 0 0 4 1134 W Example 3 Baculovirus Laspeyresia L For the preparation of a viral suspension, dead apple fruit (Laspeyresia pomonella L.) killed before and at the age of 4 (Atanassov, et. al.) with an apple fruit granule grain virus. The guns are crushed in a mortar with destilled water. After filtration and several centrifugation at 13000 rpm the pellet is dissolved in distilled water and purified by a saccharide trace (80, 60, 40, 20, 10%), and the purified granule virus granules are diluted with a little distilled water, 10 15 titer titer (granules / ml) under an electron microscope. A 10 < 6 > / ml tiger vial was then prepared. Prepared solutions were prepared from the thus prepared suspension at concentrations of 0.04-10%. For example, for 5 ml of working solution, these are 0.04% (2 μl), 0.1% (5 μl), 0.2% (10 μl), 0.4% (20 μl), 1% (50 μl) 2% (100 μl), 4% (200 μl) and 10% (500 μl). The insecticidal efficacy of the spores of the first age of apple-like worm caterpillars is tested by the method described in Example 1. The statistical processing of the data and the best results reflecting the mortality amongst the fifth day after the infestation listed in Tables 3; 3A and 3B. Table Variants of Insects in Experiment Concentration of Insecticide Test Mortality% Baculovirus Laspeyresia 60 10 500 100 - - 59 4 200 100 - 60 2 100 76 (- 60 1 50 66 Supersecret 60 0,02 100 Control - Distilled water 60 0 0 Example 4 Tobacco and blackberry extracts are prepared according to the methods described in Examples 1 and 2, then mixed in a ratio of 1: 1 but with different extracts. For example, for 5 ml of 2.5 ml of an emulsifiable solution of tobacco extract with an alkaloid content of 3,77 mg / ml and 2,5 ml of an emulsifiable solution of tea with an alkali content of 5 mg / ml The results are shown in Table 4 A. Table 4 A Variants Quantity of experiments in the assay Concentration of the insecticide Test mortality § P mg / ml% Tobacco and black extracts 60 3.77 Tobacco + 5 teaspoons 100 Extract from tobacco 60 3 , 77 95 Extract from the Americas 60 5 87 Supersect 60 0,02 100 Control - distilled water 60 0 0 5

Claims (22)

1134 U1 Example 5 A. A viral-alkaloid mixture of tobacco extract according to Example 1 was prepared containing 10 or 5 mg / ml and 100 or 200 microliters of a virion juvenile juvenile 5-10 fruit worm with a tiger 9 x 109 granules / ml for 5 ml solution. The insecticidal effect of the mixture on first apple fruit worm growers was tested by the method described in Example 1. B. A virally-alkaloid mixture of the extract of čierna according to Example 1 containing 10 or 8 mg / ml was prepared and 100 or 200 [mu] l of virus suspension 10 with a titre of 9x10 &lt; &quot; &gt; granules / ml, preferably containing the Lasoeyresia pomonella L worm granule virus for 5 ml of a solution. C. A virus-insecticidal mixture of the two extracts (tobacco and yeast) was prepared according to the method described in Example 1 containing 3,77-10 and 5-10 mg / ml extracts and a vigorous suspension of granulous virus on the apple fruit worm with a titre of 9 x 10 g / ml, in amounts of 100-200 μl per 5 ml working solution. The test of the insecticidal efficacy of the mixture was carried out according to the method described in Example 1. The test results presented in Table 5A. Table 5 A Variants Quantity of experiments in the experiment Concentration of the insecticide Mortality in the test § £% mg / ml ml% Extract from the tobacco gamma virus 60 5-10 100-200 100 Extract from the game virus virus 60 8-10 100-200 100 Ext. from tobacco and more. game virus virus 60 3,77-10 5-10 100-500 100 SuperSect 60 0,02% 100 Control - distilled water 60 0 0 GranVirus is a gyrus 9x109 granules / ml 60 100-500 100 Claims An insecticidal mixture comprising an enantiopathogenic virus comprising a total extract of Nicotianatabacum L. and / or Veratrum album L. in the following quantities in the working solution: entomopathogenesis of 4-10% vol., 0 , 5-0.65% wt. of the total extract of tobacco, extract from the strain of 0.8-1.0% by weight. and water. An insecticidal mixture as claimed in claim 1, characterized in that the endopathogenic virus is selected from the group of Baculovirus and, in particular, Baculovirus laspeyresia (Granulosa virus). An insecticidal mixture according to claim 1, characterized in that the total extract of Nicotiana tabacum L. and / or Veratrumalbum L. is pre-dissolved in a mixture of sesame oil and Tween 80® 1: 1. Insecticidal mixture according to claim 1, characterized in that the total extract is selected from alkaloid plants and in particular a total extract of Nicotianatabacum L. and / or Veratrum album L. Literature 1. Atanasov A. &amp; Stoyanova P. 1997. Investigation of the Possibilities for Enhancing the Effectiveness of Microbiological Agents for Boron Plants (Nicotiana tabacum L ). Acta Entomologica Bul-garica, 3-4, c. 67-73. 2. Atanasov A. &amp; Stoyanova P. 2001. Determination of the amount of biologically active substance in Cheat extract (Veratrum album L.), acting insecticidally against caterpillars of the lucerne fruit worm (Laspeyresia pomonella L., Lepidoptera, Tortricidae) under laboratory conditions. Acta Entomologica Bulgarica, 3-4,2001,63-67. 3. Atanasov A. &amp; Stoyanova P., 2002. Determination of the amount of biologically active tobacco substratum (Nicotiana tabacum L), active against the first age of the apple fruit worm (Laspeyresia pomonella L., Lepi-doptera, Tortricidae). In s. "Second Balkan Conference" - Quality and efficiency of the production, processing and processing of tobacco. Plovdiv, September 2002, 92-96. 4. Vassina A.N. 1970. Use of pest control systems in agriculture. Zemizdat, Sofia, 65 p. 5. Воробьева Η. N., 1976, Anthomopatogenic Viruses, "Science" Siberian Ward, Novosibirsk, 283 pp. 6. Guli VV, G. M. Ivanov, M. V. Stern-schis, 1982, Microbiological Organism of Harmful Organisms, M. Colos, 270 pp. 7. Ikonomov P., Boichinov A. &amp; НиколовП. 1941. Medicinal Plants, Sofia, Ed. Ch. Ivan Gavazov, 690 p. 8. Tarasevic L.M., 1975, Virusinssecomych, ed. "Science" Moscow, 198 p. 9. Eberle K.E., J.A.Jehle 2006, Field Resistence of Coding Moth Against Cydia Pomonella Grauulovirus (CpGV) is autosomal and incompletelydominant inherited. Journal of InvertebratePathology, 93, 201-206. 10. Sofia Katharina Kumpmann, 2002, Strategies for summer fruit tortrix moth Adoxo-phyes orana F.V. (Lepidoptera; Tortricidae) control by combining NeemAzal T / S, and productbased on Neem extracts with entomopathogenes, ph thesis. 11. Kumpmann S., J. Kienzle, C. Schulz, C.P. W; zebitz, M. Kelderer &amp; A. Trapp., 2002, Control of Adoxophyes orana F. V. R., Withentomopathogenes and NeemAzal T / S: First Approach for Optimal Combination Attractions. In: FOKO is a V (Hrsg) 10th International Conference on Cultivation Technique and Phytopathological Problems in Organic Fruit Growing and Viticulture, Weinsberg 2002; 35-42. 12. Kienzle J., Schulz C., Kumpmann S., Zebitz C. P. W., 2002, Use of NeemAzal T / S for control in apples and its combination with Bacillus thuringiensis or Granulosis for controlling Common Winter Moth or Summer Fruit TortrixMoth. In: Protection biologique and alternative pomme: verger-conservation: Colloque européen pomme.OILB / IOBC meeting Bordeaux 16-18 May 2001. 13. SU 1836005. 14. SU 2153258. 15. SU 1637653. 16. SU 2059368. 17. SU 591119. 18. US 3716634. 19. US 3639578. 20. US 3541203. 21. DE 4239222. 22. DE 4421417. 7 1134 U1 SL SL Jrb. . 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Tobacco extract - CK50 - 0.0964; fck50 = 1,023 Granulous virus - CK'50 = 0.862; fck50 = 1.024 PR = CK50: CK'50 = 8.941; fPR = 1.33 Since PR is greater than fPR, both substances differ in toxicity at P = 005; probability 95 cases per 100. <z> 00 © L / 1 O \ 4s o 4 »II S o S bJ o II II N O) O UJ Os os Os Os Os © © Os 4 © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © © -> © © © © © © © © © © © © © © © © © © © 00 00 LA 1/1 4s 4s -u LU <1 00 lu Ll 4s © 4s -J to D O m LZ1 -J Os 00 00 O OsOs Os 4s u bJ O bJ bJ 00 la Us 4s 4s 4s LU 4s oo 4s bJ Os O ru 4s 4s bJ © © © © © 4s I-bO bJ © © © © © © © © bJ © © 4s Baculovirus Laspeyresia Grape Worm Grade Table 3 Collie- Insect Mortality Concentration Insect Mortality Difference Between Insecticide Criteria Criteria in Experience Expected on Grain Fitts 4 and 6 of the experimental and expected insects (for 5 ml rabb) mortality rates-X2 pcs. % mcl lg% perforated% drilled 10 1134 W os S, W S S S O S o o m o 2 S * S3 is the N-O-to 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 X S 3 S S S S S O S O S O S LA LA LA LA LA LA LA NJ LU S P R O O S-S S S S 4 S65 C0H and 63 A5 S5 SSSO 50 S tto too X X S 3 И α α α η η st 8 8 α α S S S S S S S S S S S S S S S S S 5. 5. 5. 5. 5. 5.) σ))))) St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St St the one to IX65 S3 is a w 63 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X (X), where X 1 is the same or different and X 1 is the same or different and X 1 is X 1, ¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿ b-k- © © © © © © © © © © © © © © © © © © © 4 © tsJ bJ © bU © b © b © LU oo bU 00 © © © © © ©- © ©, 4 LU Y (B) 4 (k) - - - - - - - - - - - ‡ ‡ ‡ ‡ ‡ ‡ ‡ ‡ © © © © © © © © bJ bJ © © Mi Ml Mi © 4 4 4 * LU Cs bJ © 00 mi K) © 4 00 LU Mi © 4 © -J Ml Mi bU © - LU © -U © OO and TO oS H s H TO № 3 ss and ns ° a4 · 2 * r £ is H δ -J 3 3 OB OB wo and TO 3 Note: The calculations in ml for 5 ml of working solution.Table values for the virus refer to the 5th day WM HU w - - k kj bi 4 "- so kl Uioc oo 4 ou - - 'bJ s 4 · S * © © © © © SO 3 £ S 'and II s ts z = - 2,043 g? O 3 fa 3 3 3 H? BJ K) © O'. ‡ ‡ ‡ © © © © © © © © © © © © © © © © © © © © © © 4 © 4 © s © s o 4 4 4 4 4--00 ^ 1 (7,4- © 00 00 Axis 4b © © © © LU O © © LU O 4 © LU LU 4 © © © © © © © © © © © © LS 4 4b LU tu Is - - OO 'LU4 '71 - LJ O' OO 'Is) 4 3 3 ft) 3 "3 3 3 3 so 3 TO o 3 3 o fa a os fa 1 TO 3 3 s <3 o W ni 3 a 3 TO s <o Regression-defined regression data required for the development of bio-insecticide based on tobacco extract and pathogen-germ cellulose virus (Laspeyresia pomonella L) 12 1134 U1 coaioe ^ jO'Lft-uwM "-« OUiOOOOOOOO © O \ Os Ln <4) lo vi 4. 47 47 (> JO \ "k) ) 4 14 (7104.-4 171-L640 \ R)> - * 1740 <4 © 0000 " 0 47 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 © 0 ~ ~ © i © - © © © - © © © © © © © © - ¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¡¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿ , -0 © 70 - · K © KJ 7L KJO KJ owoseuiMOxi U> - -J-KboLu ^ KJtJioxiiJ ^ T (TiKJOK - ooo ^ 0000 l *> KJ KJ KJ KJ "" b-> kj 471 4 ^ "-. '' '' '' '' '' '' ' WU - (-) - - - - - - - - - - - - - - - - 1 W KJ © kj © - © OOKJOo '-' 1'00® 4''® Ui 4. 47 4. D * 4 »ω ω ω ωX-- 00 kj (7l 47" © 00 © 47 - ¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿ 5 l X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X Y X X 13o s ia Granulus Virus-m Glunulo w 5 The enricus ll ll ll ll ll l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l X o A in a CD M a * * * * * * * * * * * * * * * * * è è ò ò è è è è è è è è è è è è è è è è è è è è è è è è è è è è è è è è ò è è è ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò ò Office of the Republic of Bulgaria 1797 Sofia, 52 GM Dimitrov Blvd. Expert: K. Anichina No. 65365 Issued on: 40 Sep 13