Composition for Use in Controlling Aedes Aegypti

(19) &

(11) EP 1 850 666 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A01N 27/00 (2006.01) 08.08.2012 Bulletin 2012/32 (86) International application number: (21) Application number: 05769042.2 PCT/IN2005/000211

(22) Date of filing: 23.06.2005 (87) International publication number: WO 2006/090410 (31.08.2006 Gazette 2006/35)

(54) COMPOSITION FOR USE IN CONTROLLING AEDES AEGYPTI MOSQUITOES ZUSAMMENSETZUNG FÜR DIE BEKÄMPFUNG VON MOSKITOS DER ART AEDES AEGYPTI COMPOSITION DESTINEE A ETRE UTILISEE DANS LA LUTTE CONTRE LES MOUSTIQUES AEDES AEGYPTI

(84) Designated Contracting States: • CHANDEL, Kshitij FR GB Gwalior 474 002 M.P. (IN) • PARASHAR, Brahma Dutt (30) Priority: 24.02.2005 IN DE04042005 Gwalior 474 002 M.P. (IN) • VIJAYARAGHAVAN, Rajagopalan (43) Date of publication of application: Gwalior 474 002 M.P. (IN) 07.11.2007 Bulletin 2007/45 • SEKHAR, Krishnamurthy Gwalior 474 002 M.P. (IN) (73) Proprietor: Director General Defence Research & Development (74) Representative: Senior, Janet et al Organisation Abel & Imray New Delhi 110 066 (IN) 20 Red Lion Street London WC1R 4PQ (GB) (72) Inventors: • PRAKASH, Shri (56) References cited: Gwalior 474 002 M.P. (IN) • DATABASECA [Online] CHEMICAL ABSTRACTS • MENDKI, Murlidhar J. SERVICE, COLUMBUS, OHIO, US; MENDKI, M. J. Gwalior 474 002 M.P. (IN) ET AL: "Heneicosane: an oviposition-attractant • GANESAN, Kumaran pheromone of larval origin in Aedes aegypti Gwalior 474 002 M.P. (IN) mosquito" XP002362001 retrieved from STN • GOPALAN, Natarajan Database accession no. 2000:473766 & Gwalior 474 002 M.P. (IN) CURRENT SCIENCE , 78 (11), 1295-1296 CODEN: • MALHOTRA, Ramesh C. CUSCAM; ISSN: 0011-3891, 2000, Gwalior 474 002 M.P. (IN)

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 1 850 666 B1

Printed by Jouve, 75001 PARIS (FR) 1 EP 1 850 666 B1 2

Description sumably, some mosquitoes resting on vegetation at that site. Field of the invention [0010] The main drawback of this method is that it is suitable for control of mosquitoes in the field areas only [0001] The present invention relates to compositions 5 as they are highly toxic in nature thus causing an envi- for use in the control of dengue transmitting mosquito. In ronmental hazard. particular, the present invention relates to a composition [0011] Yet another drawback of this method is that for use in the control of dengue transmitting mosquito more often mosquitoes develop insecticidal resistance. comprising of an oviposition attractant to attract Aedes [0012] Still another drawback of this method is that the aegypti mosquito to lay eggs at a predetermined site and 10 adult mosquitoes which come into contact with the insec- a growth regulator, which inhibits the metamorphosis of ticides are only killed whereas the eggs/ larvae/pupae are larvae to adult mosquitoes. The present invention also unaffected by this method. relates to processes for preparation of such composi- [0013] Still another drawback of this method is that tions. The present invention also relates to methods for non-target organisms are also killed along with mosqui- controlling Aedes aegypti mosquito and use of the com- 15 toes such as aquatic fish, etc. Thus it is not environment positions of the present invention for controlling Aedes and eco-friendly. aegypti mosquitoes. [0014] Another method known in the art of control of mosquitoes involves larval management. This includes Description of the prior art the use of chemical larvicides, microbial insecticides (Ba- 20 cillus thuringiensis B thuringiensis israeliensis), larvicid- [0002] Mosquitoes are medically important vectors, al oils, juvenile hormone (methoprene), and mosquito both in number of diseases they transmit and the mag- predator fish to control mosquito larvae. nitude of health problems world-wide. About 400 species [0015] The main disadvantage of these methods is that of mosquitoes can spread disease to humans. These in- they are suitable for use in ponds, canals, irrigated fields clude yellow fever and dengue hemorrhagic fever for25 and some other freshwater sources for mosquito larval which the mosquitoes, Aedes aegypti and Aedes al- control only. bopictus are the important vectors. According to the [0016] Another disadvantage of these methods is that World Health Organization, at least 700 million people closer surveillance is required to find out the larval breed- each year are infected with disease by mosquitoes. ing sites for the application of larvicides. [0003] Dengue fever is a serious disease of Asia and 30 [0017] Still another disadvantage of microbial control Africa. In India, public is aware of dengue haemorrhagic and mosquito predator fish control methods are that they fever due to the epidemics in 1996 particularly in Delhi cannot be used in a container breeding mosquitoes like and surrounding regions. Classic dengue, known for its Aedes aegypti. low mortality but very uncomfortable symptoms, has be- [0018] Further another disadvantage of these methods come more serious, both in frequency and mortality, in 35 is that the mosquitoes develop resistance to microbial recent years. Aedes aegypti mosquito is the most prom- agents. inent vector of dengue throughout the world. World wide [0019] Yet another disadvantage of microbial larvicide at present there is no dengue vaccine. Therefore, mos- is that it degrades faster in presence of sun light. quito control measures are the only remedy to this prob- [0020] Another disadvantage of these methods, which lem. 40 are based on larvicides, is that they are not economical [0004] A variety of methods have been practised in the to use for the control of dengue transmitting mosquitoes. art in an attempt to control Aedes mosquitoes. [0021] Another disadvantage of the method using [0005] One of the methods used for the control of mos- mosquito predator fish to eliminate mosquito larvae is quito involves attract and kill devices which uses different that the life- period of the mosquito predator fish depends combinations such as ultraviolet light, Carbondioxide, 45 on the specific characteristics of the water like salinity. and 1-octen-3-ol to attract mosquitoes. Maintenance of such conditions poses limitations for the [0006] The main disadvantage of this method is that practical application of this method. these devices are not efficient. [0022] Still another disadvantage of this method is that [0007] Yet another disadvantage of this method is that it cannot be used for monitoring the vector population. these devices are not suitable for control of mosquitoes 50 [0023] Yet another disadvantage of these methods is in the field conditions. that they are not cost effective. [0008] Still another disadvantage of this method is that [0024] Thus, there is an urgent need for a new mos- it is an expensive method for its public use. quito control composition and method, which is simple [0009] Another method known in the art for controlling to use, cost-effective, and environment and eco- friendly. mosquitoes is the use of insecticides such as organo- 55 phosphorus pesticides (malathion or fenitrothion), car- Objects of the present invention bamates and synthetic pyrethroids by residual application or fogging to kill flying adult mosquitoes and, pre- [0025] It is therefore, one of the main objects of the

2 3 EP 1 850 666 B1 4 present invention to provide a composition for the control [0035] The present invention also provides a ‘lure and of dengue transmitting mosquito Aedes aegypt, which kill’ composition for luring Aedes aegypti mosquitoes to overcomes the disadvantages of the prior art. lay their eggs in a target site and killing the mosquito [0026] Another object of the present invention is to pro- larvae or regulating the growth of mosquito larvae, which vide an environment and eco- friendly composition for the 5 comprises (a) an effective amount of n- heneicosane dis- control of dengue transmitting mosquito Aedes aegypt, solved in an organic solvent, and (b) an effective amount which does not contain any toxic chemical. of an insect growth regulator dissolved in an organic sol- [0027] Another object of the present invention is to pro- vent. vide a composition, which can be used easily in ovitraps, [0036] The present invention further provides a ’lure coolers, overhead tanks and other dengue transmitting 10 and kill’ composition for luring Aedes aegypti mosquitoes mosquito breeding aquatic habitats. to lay their eggs in a target site and killing the mosquito [0028] Still another object of the present invention is larvae or regulating the growth of mosquito larvae, which to provide a novel chemical composition, which attracts comprises an effective amount of n-heneicosane dis- dengue transmitting mosquitoes to lay eggs at a given solved in a suitable organic solvent, and an effective site and control mosquito population by blocking the lar- 15 amount of an insecticide. val growth to adults. [0037] In these embodiments, the present invention [0029] Yet another object of the present invention is to utilises a composition for luring Aedes aegypti mosqui- provide a novel chemical composition which can be used toes to lay their eggs in a target site which comprises of for the control of breeding of dengue transmitting mos- an effective amount of n-heneicosame in a suitable or- quitoes in indoor as well as outdoor applications. 20 ganic solvent. [0030] Still another object of the present invention is [0038] In a preferred feature, said solvent is selected to provide a novel chemical composition for the control from as petroleum ether, n-hepentane, n-hexane, meth- of dengue transmitting mosquito Aedes aegypt, which anol and dichloromethane. does not affect the other beneficial insects in the same [0039] In a preferred feature, the ratio ofhenei- n- aquatic habitat. 25 cosane to said solvent is, preferably, from 1:20 to 1:70. [0031] Yet another object of the present invention is to [0040] A lure and kill composition of the invention fur- provide a novel chemical composition for nearly 100% ther comprises an effective amount of an insect growth control of dengue transmitting mosquito Aedes aegypti, regulator or insecticide. within 2 days of use. [0041] In a preferred feature of the invention the insec- 30 ticide is a larvicide. Summary of the invention [0042] In a preferred feature of the invention, the insect growth regulator is selected from diflubenzuron, neema- [0032] The present invention is based on the unex- zal and triflumuron. pected finding that an oviposition attractant pheromone [0043] In a preferred feature of the invention, said in- of the dengue transmitting mosquito, Aedes aegypti ,in 35 sect growth regulator comprises 100 to 120 mg of dif- combination with a suitable growth regulator in proper lubenzuron in 500 ml of an organic solvent selected from composition prevents the development of Aedes aegypti acetone and dichloromethane. at larval stage itself, thus controlling the mosquito popu- [0044] Preferably, said n-heneicosane concentration lation by ’lure and kill technique’. is 8 to 20 g, more preferably 10 to 12 g, per litre of the [0033] Accordingly the present invention provides a 40 composition. ‘lure and kill’ composition for luring Aedes aegypti mos- [0045] The present invention also provides a process quitoes to lay their eggs in a target site and killing the for the preparation of a composition for the control of mosquito larvae or regulating the growth of mosquito lar- dengue transmitting mosquito having lure and kill prop- vae, comprising of an oviposition attractant pheromone erty comprising of the steps of: n-heneicosane and an effective amount of an insect45 growth regulator in an inert carrier. The said chemical (a) preparing oviposition attractant solution by dis- composition controls dengue transmitting mosquito pop- solving n-heneicosane in an organic solvent; ulation by attracting the females to lay eggs in water (b) preparing insect regulator solution by dissolving where it is applied and stops further development of lar- an insect growth regulator in an organic solvent; vae to adults. This chemical composition can be applied 50 (c) mixing the above two solutions thoroughly. to any Aedes aegypti mosquito breeding in aquatic habitat. [0046] In a preferred feature, the ratio ofhenei- n- [0034] The composition can be used in different for- cosane to said solvent is, preferably, from 1:20 to 1:70. mulations such as spray, tablets and lotions and it kills [0047] In a preferred feature, the insect growth regu- the larvae within two days after hatching from the egg. 55 lator comprises 100-120 mg of diflubenzuron in 500 ml The mortality rate is 100%. This method of Aedes aegypti of an organic solvent selected from acetone and dichlo- mosquito control is simple, cost-effective, efficient and romethane. eco-friendly. [0048] In a preferred feature, the process comprises

3 5 EP 1 850 666 B1 6 preparing oviposition attractant solution by dissolving 8 gypti mosquitoes tend to lay eggs in stagnant water to 20 g of n-heneicosane, preferably 10 - 12 g, in 500 ml where eggs of the same species of mosquitoes had been of organic solvents like n-pentane, n-hexane or dichlo- laid previously. This led the inventors to analyse various romethane, preparing insect regulator solution by dis- samples of water which in turn led to the finding that water solving 800 - 2000 mg of insect growth regulator like di- 5 in which eggs had previously been laid by Aedes aegypti flubenzuron, preferably 100 - 120 mg, in 500 ml of organic mosquitoes and which had hatched into larval stage con- solvents like acetone or dichloromethane and mixing the tained various semio-chemicals. Analysis of these sem- above two solutions thoroughly. io-chemicals identified them as heneicosane, nona- [0049] The present invention utilises n-heneicosane cosane, octadecane, isopropyl myristate and docosane. for luring Aedes aegypti mosquitoes to lay their eggs in 10 Of these five chemicals, heneicosane was found to be a target site, and provides a method for the control of most effective oviposition attractant which chemically "in- Aedes aegypti mosquitoes which comprises luring the vites" the mosquitoes to lay their eggs in the same place mosquitoes to lay their eggs in a target site and killing again and again. When stagnant water in which Aedes the resulting larvae or regulating the growth of mosquito aegypti mosquitoes had bred earlier was left untouched larvae, which method comprises treating the site with (a) 15 the incidence of further egg laying in the same water and an effective amount of n-heneicosane dissolved in an thriving of larval population increased. However, when organic solvent and with (b) (i) an effective amount of an such water was replaced by fresh water the incidence of insect growth regulator dissolved in an organic solvent, egg laying decreased considerably and thereafter, grad- or (ii) an effective amount of an insecticide. ually increased over a period of time. However, when [0050] According to the present invention a method for 20 stagnant water was kept alongside water containing he- the control of Aedes aegypti mosquitoes may comprise neicosane, the entire egg laying took place only in water treating the target site with an effective amount of a com- containing heneicosane thereby clearly establishing that position of the invention. the Aedes aegypti mosquitoes were attracted to water [0051] In one embodiment, a method for the control of containing heneicosane for the purposes of laying their Aedes aegypti mosquitoes comprises treating said target 25 eggs. Thus, the present invention makes it possible to site with an effective amount of n-heneicosane dissolved lure Aedes aegypti mosquitoes to lay their eggs only at in a suitable organic solvent, said composition further specific chosen sites making it possible to thereafter to comprising an effective amount of an insect growth reg- destroy the entire larval population and control further ulator or insecticide. breeding of the mosquitoes. This has a tremendous in- 30 dustrial application. It minimises the use of insecticides Detailed Description and therefore, wastage thereof, since in accordance with the present invention, the insecticides are required to be [0052] The present invention relates to the control of used only at specific target sites instead of over a large Aedes aegypti mosquitoes based on "lure and kill" tech- area. It also ensures that the insects do not escape. nique. These mosquitoes are lured chemically to lay eggs 35 Spraying of insecticides over a large area offers tremen- only at a specific site. When the eggs are hatched, the dous opportunities to an adult mosquito to escape. On entire larvae population can be destroyed. A composition the contrary, present invention ensures that the entire comprising n-heneicosane used in the present invention larval population is congregated at a specific site where ensures that the eggs are laid only in a specific site and 100% killing thereof can be achieved. not anywhere else. 40 [0056] The present invention also relates to a process [0053] The present invention is based on the unex- for the preparation of the composition of the present in- pected finding that heneicosane, a C- 21 hydrocarbon, is vention. In the first aspect, the invention relates to prep- capable of acting as a most effective oviposition attract- aration and isolation of heneicosane. In this embodiment, ant. Thus, any standing water containing a dose of he- Aedes aegypti mosquitoes are bred and allowed to lay neicosane will attract and lure Aedes aegypti mosquitoes 45 eggs in stagnant water. Once the eggs are hatched and to lay their eggs only in such water and no other. Subse- during the larval stage thereof, the water is collected. quently, either during the larval stage or just before the This water is referred to as larval conditioned water eggs are hatched, an effective dosage of a conventional (LCW). The semio-chemicals are thereafter extracted insecticide may be added to completely destroy the lar- from the LCW. Conventional organic solvents such as vae. 50 petroleum, ether, hexane, dichloromethane, or methanol [0054] In an alternative embodiment, heneicosane are used for the extraction of the semio-chemicals. The may be synergistically premixed with an insecticide so most preferred solvent is hexane. The extracted solution that the composition itself acts as a "lure and kill" com- comprised of the following five somio-chemicals: henei- position. cosane, nonacosane, octadecane, isopropyl myristate [0055] The present invention also relates to a process 55 and docosane. From the mixture eof such five semio- for the preparation of compositions for use in lure and/or chemicals, heneicosane is separated. killing of Aedes aegypti mosquitoes. The present inven- [0057] A composition comprising or containing henei- tion is based on the unexpected finding that Aedes ae- cosane is sprayed into water referred to herein as target

4 7 EP 1 850 666 B1 8 site. The composition may include any insecticide. In the by "lure and kill" technique. next few days, there is a tremendous Incidence of egg [0065] The .above chemical composition can be used laying activity by Aedes aegypti mosquitoes at the target in different formulations such as spray, tablets and lo- site.The composition may already contain a conventional tions. Extensive field trials with this chemical composition insecticide, which is effective against Aedes aegypti lar- 5 conducted after adequate precaution of confidentiality in- vas. Alternatively, the target site is treated with a con- dicated that larvae after hatching from the eggs die within ventional insecticide or insect growth regulator such as 2 days and the mortality rate is 100%. The water in ovit- diflubenzuron, neomazal, triflumuron. Diflubenzuron is raps containing this chemical composition is found to be the preferred insect growth regulator. effective for 7 to 10 days to attract the Aedes aegypti to [0058] In a preferred embodiment, solutions of ovipo- 10 lay eggs and kill the larvae a second time after hatching sition attractant containing heneicosane and insect from the eggs relaid in the same water. This method of growth regulator are prepared and kept separately. mosquito control is simple, cost effective, efficient and These two compositions are subsequently mixed in a pre- eco-friendly. determined ratio, preferably at a ratio of 1: 1 The two com- [0066] In a preferred embodiment, the process for the positions may be mixed either in advance or just prior to 15 preparation of the composition of the present invention use. comprises comprising of the steps of: [0059] The composition of the present invention has been found to be extremely effective in "lure and kill" (a) preparing oviposition attractant solution by dis- technique against Aedes aegypti mosquitoes. solving 8 to 20 g of n- heneicosane preferably 10-12 [0060] In a preferred embodiment, the process in ac- 20 g in 500 ml of organic solvents like pentane,n- n- cordance with the present invention for the preparation hexane or dichloromethane. of new chemical composition for the control of dengue (b) preparing insect regulator solution by dissolving transmitting mosquitoes comprises of following steps: 800-2000 mg of insect growth regulator like diflubenzuron preferably 100-120 mg in 500 ml of organic Step (a): Preparation of oviposition attractant solution: 25 solvents like acetone or dichloromethane.

[0061] Aedes aegypti mosquitoes are bred and the wa- [0067] The present invention will now be described in ter is collected for isolation where the mosquitoes are at greater detail with the following examples. It be under- larval stage. This water is called as larval conditioned stood by a person skilled in the art that the examples are water (LCW). The semio-chemicals are extracted from 30 for illustrative purposes only and various modifications the LCW by organic solvents such as petroleum ether, of the present invention are possible without departing hexane, dichloromethane or methanol preferably hex- from the spirit and scope thereof ane. The extractant is analysed by various spectral tech- niques like IR, NMR and GCMS and the five semio-chem- Example 1 icals identified are heneicosane, nonacosane, octade- 35 cane, isopropyl myristate and decosane. Amongst these The following examples illustrate the process for prepar - heneicosane (C 21 hydrocarbon) is found to be the most ing the composition of the present invention effective oviposition attractant. [0062] In a stoppered bottle, 1.6 to 2% w/v preferably [0068] To a 1 Lstoppered bottle, 250ml ofor n-pentane 1.0-1.2% w/v of n-heneicosane is dissolved completely 40 (HPLC grade) is taken. To this 13 g of n-heneicosane g in n-pentane or n-hexane HPLC grade preferably n-hex- is added and the flask is shaken well to dissolve it com- ane. pletely. [0069] 250 ml of acetone HPLC grade is taken in an- Step (b): Preparation of insect growth regulator solution: other 500 ml of stoppered bottle. To this 100 mg of dif- 45 lubenzuron is added and the flask is shaken well to dis- [0063] In a stoppered bottle, 0.16 to 0.2% w/v, prefer- solve it completely. This is transferred to the bottle con- ably, 0.1 to 0.12% w/v of insect growth regulator like di- taining the heneicosane solution and the mixture is shak- flubenzuron, neemazal, triflumuron preferably difluben- en thoroughly. This composition is ready for use for the zuron is dissolved completely in acetone HPLC grade. control of dengue transmitting mosquito breeding. 50 Step (c): Preparation oft he chemical compositionhaving Example 2 lure and kill property: [0070] To a L stoppered bottle, 250 ml of n-hexane [0064] The oviposition attractant solution prepared in HPLC grade is taken. To this 10 g of n- heneicosane g is step (a) and insect growth regulator solution prepared in 55 added and the flask is shaken well to dissolve it com- step (b) are taken in a ratio of 11: and the mixture is pletely. shaken thoroughly. This composition can readily be used [0071] 250 ml of acetone HPLC grade is taken in an- to control the dengue transmitting mosquitoes breeding other 500 ml of stoppered bottle. To this 100 mg of dif-

5 9 EP 1 850 666 B1 10 lubenzuron is added and the flask is shaken well to dis- EXAMPLE 6 solve it completely. This is transferred to the bottle containing the heneicosane solution and the mixture is shak- [0077] Example Nos. 4 and 5 were repeated with he- en thoroughly. This composition is ready for use for the neicosane dissolved in n-pentane. The results were ex- control of dengue transmitting mosquito breeding. 5 actly the same.

Example 3 EXAMPLE 7

[0072] To a L stoppered bottle, 250 ml of or n- pentane [0078] The above example was repeated with varying HPLC grade is taken. To this 20 g of n-heneicosane is 10 concentrations of heneicosane in the solvent. The con- added and the flask is shaken well to dissolve it com- centration of 1.0 to 2% w/v was found to be most effective. pletely. [0073] 250 of acetone HPLC grade is taken in another 500 ml of stoppered bottle. To this 150 mg of difluben- Claims zuron is added and the flask is shaken well to dissolve it 15 completely. This is transferred to the bottle containing 1. A ’lure and kill’ composition for luring Aedes aegypti the heneicosane solution and the mixture is shaken thor- mosquitoes to lay their eggs in a target site and killing oughly. This composition is ready for use for the control the mosquito larvae or regulating the growth of mos- of dengue transmitting mosquito breeding. quito larvae, which comprises (a) an effective 20 amount of n-heneicosane dissolved in an organic EXAMPLE 4 solvent, and (b) an effective amount of an insect growth regulator dissolved in an organic solvent. The following examples illustrate the effectiveness of the "luring" effect. 2. A composition as claimed in claim 1, wherein the 25 insect growth regulator is selected from difluben- [0074] The following experiment was conducted under zuron, neemazal and triflumuron. field conditions in a specially cordoned area to which public did not have any access. Fifteen open tanks with water 3. A composition as claimed in claim 1 or claim 2, filled to the top were kept in the open. All the tanks were wherein the concentration of n-heneicosane is from made of same material, were of the same size and con- 30 8 to 20 g per litre of the composition. tained same amount of water from the same source. The tanks were numbered consecutively and allowed to re- 4. A composition as claimed in claim 3, wherein the n- main for a week. After fifteen days all the tanks were heneicosane concentration is 10-12 g per litre of the examined and it was found that all the tanks contained composition. nearly the same amount of larval population ofAedes 35 aegypti mosquitoes. This establishes that the mosqui- 5. A composition as claimed in any one of claims 1 to toes did not have any preference for any particular water 4, wherein component (a) comprises n-heneicosane tank. dissolved in petroleum ether, n-pentane, n-hexane, [0075] All the fifteen tanks were drained of water and methanol or dichloromethane. thoroughly cleaned. Subsequently, they were filled with 40 water from the same source. In tank No. 1, a predeter- 6. A composition as claimed in claim 3, wherein the mined amount of a solution of heneicosane in n-hexane concentrations of n-heneicosane and insect growth (1% w/v) was added. After fifteen days, water from each regulator correspond to those of a composition pre- tank was analysed. It was found that only tank No. 1 pared by mixing (a) 8 to 20 g of n-heneicosane in containing the heneicosane contained mosquito larvae, 45 500 ml of an organic solvent with (b) 800 to 2000 mg thereby establishing thatAedes aegypti mosquitoes of insect growth regulator in 500ml of an organic sol- were clearly attracted and lured to lay eggs in water con- vent. taining heneicosane. 7. A composition as claimed in claim 6, wherein the n- EXAMPLE 5 50 heneicosane is pre-dissolved in n-pentane, n-hexane or dichoromethane, and mixed with insect [0076] Example No. 4 was repeated 14 times with he- growth regulator solution. neicosane being added in tank Nos. 2 to 15 consequen- tially. It was found that each time the tank containing he- 8. A composition as claimed in claim 6 or claim 7, neicosane attracted Aedes aegypti mosquitoes to lay 55 wherein the insect growth regulator is diflubenzuron. their eggs therein. 9. A composition as claimed in any one of claims 6 to 8, wherein the insect growth regulator is pre-dis-

6 11 EP 1 850 666 B1 12

solved in acetone or dichloromethane, and mixed neemazal and triflumuron. with n-heneicosane solution. 20. A process as claimed in any one of claims 17 to 19, 10. A composition as claimed in any one of claims 7 to wherein the concentration of n-heneicosane is from 9, which contains 10 to 12 g of n-heneicosane per 5 8 to 20 g per litre of the composition. litre of composition. 21. A process as claimed in claim 20, wherein said n- 11. A composition as claimed in claim 1 or claim 2, heneicosane concentration is 10-12 g per litre of the wherein the concentrations of n- heneicosane and in- composition. sect growth regulator correspond to those of a com- 10 position prepared by mixing, in a ratio of 1:1, 1.0 to 22. A process as claimed in any one of claims 17 to 21, 2 % w/v of n-heneicosane in n- pentane or n-hexane wherein in step (a) the n-heneicosane is dissolved with 0.1 to 0.2 % w/v of an insect growth regulator in petroleum ether, n-pentane, n-hexane, methanol in acetone. or dichloromethane. 15 12. A ’lure and kill’ composition for luring Aedes aegypti 23. A process as claimed in claim 20, which comprises mosquitoes to lay their eggs in a target site and killing the mosquito larvae or regulating the growth of mos- (a) preparing oviposition attractant solution by quito larvae, which comprises dissolving 8 to 20 g of n-heneicosane in 500 ml 20 of organic solvent, a) an effective amount of n-heneicosane, and (b) preparing insect regulator solution by dis- b) an effective amount of an insect growth reg- solving 800-2000 mg of insect growth regulator ulator, in 500 ml of organic solvent, and in an inert carrier. (c) mixing the above two solutions thoroughly. 25 13. A composition as claimed in claim 12, wherein the 24. A process as claimed in any one of claims 17 to 19, insect growth regulator is selected from difluben- which comprises mixing, in a ratio of 1: 1, 1.0 to 2 % zuron, neemazal and triflumuron w/v of n-heneicosane in n-pentane or n- hexane with 0.1 to 0.2 % w/v of an insect growth regulator in ac- 14. A composition as claimed in claim 12 or claim 13, 30 etone. which is in the form of a spray. 25. A method for the control of Aedes aegypti mosqui- 15. A ’lure and kill’ composition for luring Aedes aegypti toes which comprises luring the mosquitoes to lay mosquitoes to lay their eggs in a target site and killing eggs in a target site and killing the resulting larvae the mosquito larvae or regulating the growth of mos- 35 or regulating the growth of mosquito larvae by treat- quito larvae, which comprises an effective amount ing the site with (a) an effective amount of n-henei- of n-heneicosane dissolved in a suitable organic sol- cosane dissolved in an organic solvent, and with (b) vent, and an effective amount of an insecticide. (i) an effective amount of an insect growth regulator dissolved in an organic solvent, or (ii) an effective 16. A composition as claimed in claim 15, wherein the 40 amount of an insecticide. insecticide is a larvicide. 26. A method as claimed in claim 25, wherein the treat- 17. A process for the preparation of a composition for ment is carried out with a composition according to the control of dengue transmitting mosquito Aedes any one of claims 2 to 11. aegypti, comprising of the steps of: 45 27. A method as claimed in claim 25 or claim 26, wherein (a) preparing oviposition attractant solution by components (a) and (bi) are mixed in advance or just dissolving n-heneicosane in an organic solvent; prior to use. (b) preparing insect regulator solution by dissolving an insect growth regulator in an organic 50 28. A method for the control of Aedes aegypti mosqui- solvent; and toes which comprises luring the mosquitoes to lay (c) mixing the above two solutions thoroughly. eggs in a target site and killing the resulting larvae or regulating the growth of mosquito larvae by treat- 18. A process as claimed in claims 17, wherein the insect ing the site with a composition as claimed in any one growth regulator is a larvicide. 55 of claims 12 to 16.

19. A process as claimed in claim 18, wherein the insect growth regulator is selected from diflubenzuron,

7 13 EP 1 850 666 B1 14

Patentansprüche 11. Zusammensetzung, wie in Anspruch 1 oder in An- spruch 2 beansprucht, wobei die Konzentrationen 1. Zusammensetzung vom Typ ’Anlocken und Vernich- an n-Heneicosan und Wachstumsregler für Insekten ten’ (’Lure and Kill’), um Stechmücken der Art Aedes denen einer Zusammensetzung entsprechen, die aegypti anzulocken, ihre Eier an einen Zielort zu le- 5 hergestellt wird, indem 1,0 bis 2% w/v n- Heneicosan gen, und um die Stechmückenlarven zu töten oder in n-Pentan oder n- Hexan mit 0,1 bis 0,2% w/v eines das Wachstum der Stechmückenlarven zu regeln, Wachstumsreglers fürInsekten in Aceton im Verhält- welche (a) eine wirksame Menge an n- Heneicosan, nis 1:1 gemischt werden. das in einem organischen Lösungsmittel gelöst ist, und (b) eine wirksame Menge eines Wachstumsreg- 10 12. Zusammensetzung vom Typ ’Anlocken und Vernich- lers für Insekten, der in einem organischen Lösungs- ten’ (’Lure and Kill’), um Stechmücken der Art Aedes mittel gelöst ist, umfasst. aegypti anzulocken, ihre Eier an einen Zielort zu legen, und um die Stechmückenlarven zu töten oder 2. Zusammensetzung, wie in Anspruch 1 beansprucht, das Wachstum der Stechmückenlarven zu regeln, wobei der Wachstumsregler für Insekten aus Diflu- 15 welche benzuron, Neemazal und Triflumuron ausgewählt wird. (a) eine wirksame Menge an n- Heneicosan, und (b) eine wirksame Menge eines Wachstumsreg- 3. Zusammensetzung, wie in Anspruch 1 oder in An- lers für Insekten in einem inerten Träger um- spruch 2 beansprucht, wobei die Konzentration an 20 fasst. n-Heneicosan von 8 bis 20 g pro Liter der Zusam- mensetzung beträgt. 13. Zusammensetzung, wie in Anspruch 12 beansprucht, wobei der Wachstumsregler aus Difluben- 4. Zusammensetzung, wie in Anspruch 3 beansprucht, zuron, Neemazal und Triflumuron, ausgewählt wird. wobei die n-Heneicosan-Konzentration 10 bis 12 g 25 pro Liter der Zusammensetzung beträgt. 14. Zusammensetzung, wie in Anspruch 12 oder in An- spruch 13 beansprucht, welche in Form eines 5. Zusammensetzung, wie in einem der Ansprüche 1 Sprays ist. bis 4 beansprucht, wobei die Komponente (a) n- Heneicosan umfasst, das in Petrolether, n-Pentan, 30 15. Zusammensetzung vom Typ ’Anlocken und Vernich- n-Hexan, Methanol oder Dichlormethan gelöst ist. ten’ (’Lure and Kill’), um Stechmücken der Art Aedes aegypti anzulocken, ihre Eier an einen Zielort zu le- 6. Zusammensetzung, wie in Anspruch 3 beansprucht, gen, und um die Stechmückenlarven zu töten oder wobei die Konzentrationen an n-Heneicosan und das Wachstum der Stechmückenlarven zu regeln, Wachstumsregler für Insekten denen einer Zusam- 35 welche eine wirksame Menge an n- Heneicosan, das mensetzung entsprechen, die hergestellt wird, in- in einem geeigneten organischen Lösungsmittel ge- dem (a) 8 bis 20 g n-Heneicosan in 500 ml eines löst ist, und eine wirksame Menge eines Insektizides organischen Lösungsmittels mit (b) 800 bis 2000 mg umfasst. Wachstumsregler für Insekten in 500 ml eines organischen Lösungsmittels gemischt werden. 40 16. Zusammensetzung, wie in Anspruch 15 beansprucht, wobei das Insektizid ein Larvizid ist. 7. Zusammensetzung, wie in Anspruch 6 beansprucht, wobei das n-Heneicosan in n- Pentan, n-Hexan oder 17. Verfahren zur Herstellung einer Zusammensetzung Dichlormethan vorab gelöst ist, und mit einer Lösung für die Kontrolle von Dengue-Fieber-übertragenden von Wachstumsregler für Insekten gemischt ist. 45 Stechmücken der Art Aedes aegypti, umfassend die folgenden Schritte: 8. Zusammensetzung, wie in Anspruch 6 oder in An- spruch 7 beansprucht, wobei der Wachstumsregler (a) Herstellen einer Lösung eines Lockmittels für Insekten Diflubenzuron ist. für die Eiablage, indem n-Heneicosan in einem 50 organischen Lösungsmittel gelöst wird; 9. Zusammensetzung, wie in einem der Ansprüche 6 (b) Herstellen einer Lösung eines Insektenreg- bis 8 beansprucht, wobei der Wachstumsregler für lers, indem ein Wachstumsregler für Insekten in Insekten in Aceton oder Dichlormethan vorab gelöst einem organischen Lösungsmittel gelöst wird; ist, und mit n-Heneicosan-Lösung gemischt ist. und 55 (c) gründliches Mischen der beiden obigen Lö- 10. Zusammensetzung, wie in einem der Ansprüche 7 sungen. bis 9 beansprucht, welche 10 bis 12 g n- Heneicosan pro Liter der Zusammensetzung enthält. 18. Verfahren, wie in Anspruch 17 beansprucht, wobei

8 15 EP 1 850 666 B1 16

der Wachstumsregler für Insekten ein Larvizid ist. im Voraus oder direkt vor Verwendung gemischt werden. 19. Verfahren, wie in Anspruch 18 beansprucht, wobei der Wachstumsregler für Insekten aus Diflubenzu- 28. Verfahren für die Kontrolle von Stechmücken der Art ron, Neemazal und Triflumuron ausgewählt wird. 5 Aedes aegypti, welches das Anlocken der Stech- mücken, damit sie Eier an einen Zielort legen, und 20. Verfahren, wie in einem der Ansprüche 17 bis 19 das Töten der resultierenden Larven oder das Re- beansprucht, wobei die Konzentration an n-Henei- geln des Wachstums der Stechmückenlarven durch cosan von 8 bis 20 g pro Liter der Zusammensetzung Behandeln des Ortes mit einer Zusammensetzung, beträgt. 10 wie in einem der Ansprüche 12 bis 16 beansprucht, umfasst. 21. Verfahren, wie in Anspruch 20 beansprucht, wobei die n-Heneicosan-Konzentration 10 bis 12 g pro Liter der Zusammensetzung beträgt. Revendications 15 22. Verfahren, wie in einem der Ansprüche 17 bis 21 1. Une composition "lure and kill" ("attirer et tuer") pour beansprucht, wobei in Schritt (a) das n- Heneicosan attirer les moustiques Aedes aegypti de manière à in Petrolether, n-Pentan, n-Hexan, Methanol oder ce qu’ils pondent leurs oeufs dans un site cible et Dichlormethan gelöst wird. pour tuer les larves de moustiques ou réguler la 20 croissance des larves de moustiques, qui comprend 23. Verfahren, wie in Anspruch20 beansprucht, welches (a) une quantité efficace de n- heneicosane dissoute umfasst: dans un solvant organique, et (b) une quantité efficace d’un régulateur de croissance d’insectes dis- (a) Herstellen einer Lösung eines Lockmittels soute dans un solvant organique. für die Eiablage, indem 8 bis 20 g n- Heneicosan 25 in 500 ml eines organischen Lösungsmittels ge- 2. Une composition selon la revendication 1, dans la- löst werden, quelle le régulateur de croissance d’insectes est sé- (b) Herstellen einer Lösung eines Insektenreg- lectionné parmi le diflubenzuron, le neemazal et le lers, indem 800 bis 2000 mg Wachstumsregler triflumuron. für Insekten in 500 ml eines organischen Lö- 30 sungsmittels gelöst werden, und 3. Une composition selon la revendication 1 ou la re- (c) gründliches Mischen der beiden obigen Lö- vendication 2, dans laquelle la concentration de n- sungen. heneicosane est de 8 à 20 g par litre de composition.

24. Verfahren, wie in einem der Ansprüche 17 bis 19 35 4. Une composition selon la revendication 3, dans la- beansprucht, welches ein Mischen von 1,0 bis 2% quelle la concentration de n-heneicosane est de 10 w/v n-Heneicosan in n- Pentan oder n-Hexan mit 0,1 à 12 g par litre de composition. bis 0,2% w/v eines Wachstumsreglers für Insekten in Aceton in einem Verhältnis von 1:1 umfasst. 5. Une composition selon l’une quelconque des reven- 40 dications 1 à 4, caractérisé en ce que le composant 25. Verfahren für die Kontrolle von Stechmücken der Art (a) comprend du n-heneicosane dissous dans de Aedes aegypti, welches das Anlocken der Stech- l’éther de pétrole, du n-pentane, du n-hexane, du mücken, damit sie Eier an einen Zielort legen, und méthanol ou du dichlorométhane. das Töten der resultierenden Larven oder das Re- geln des Wachstums der Stechmückenlarven durch 45 6. Une composition selon la revendication 3, dans la- Behandeln des Ortes mit (a) einer wirksamen Menge quelle les concentrations en régulateur de croissan- an n-Heneicosan, das in einem organischen Lö- ce d’insectes et en n-heneicosane correspondent à sungsmittel gelöst ist, und mit (b) (i) einer wirksamen celles d’une composition préparée en mélangeant Menge eines Wachstumsreglers für Insekten, der in (a) de 8 à 20 g de n-heneicosane dans 500 ml d’un einem organischen Lösungsmittel gelöst ist, oder (ii) 50 solvant organique avec (b) de 800 à 2000 mg de einer wirksamen Menge eines Insektizides umfasst. régulateur de croissance d’insectes dans 500 ml d’un solvant organique. 26. Verfahren, wie in Anspruch 25 beansprucht, wobei die Behandlung mit einer Zusammensetzung nach 7. Une composition selon la revendication 6, dans la- einem der Ansprüche 2 bis 11 ausgeführt wird. 55 quelle le n-heneicosane est préalablement dissous dans du n-pentane, du n-hexane ou du dichloromé- 27. Verfahren, wie in Anspruch 25 oder in Anspruch 26 thane, et est mélangé avec une solution de régula- beansprucht, wobei die Komponenten (a) und (b)(i) teur de croissance d’insectes.

9 17 EP 1 850 666 B1 18

8. Une composition selon la revendication 6 ou la re- (a) préparer une solution attractive pour la ponte vendication 7, dans laquelle le régulateur de crois- des oeufs en dissolvant du n- heneicosane dans sance d’insectes est le diflubenzuron. un solvant organique ; (b) préparer une solution de régulation d’insec- 9. Une composition selon l’une quelconque des reven- 5 tes en dissolvant un régulateur de croissance dications 6 à 8, dans laquelle le régulateur de crois- d’insectes dans un solvant organique ; et sance d’insectes est pré-dissous dans de l’acétone (c) mélanger intimement les deux solutions ci- ou du dichlorométhane, et est mélangé avec une dessus. solution de n-heneicosane. 10 18. Un procédé tel que revendiqué dans la revendication 10. Une composition selon l’une quelconque des reven- 17, dans lequel le régulateur de croissance d’insec- dications 7 à 9, qui contient 10 à 12 g de n-heneico- tes est un larvicide. sane par litre de composition. 19. Un procédé tel que revendiqué dans la revendication 11. Une composition selon la revendication 1 ou la re- 15 18, dans lequel le régulateur de croissance d’insec- vendication 2, dans laquelle les concentrations en tes est sélectionné parmi le diflubenzuron, le nee- n-heneicosane et en régulateur de croissance d’in- mazal et le triflumuron. sectes correspondent à celles d’une composition préparée en mélangeant, dans un rapport de 1:1, de 20. Un procédé tel que revendiqué dans l’une des re- 1,0 à 2% p/v de n-heneicosane dans du n-pentane 20 vendications 17 à 19, dans lequel la concentration ou du n-hexane avec de 0,1 à 0,2% p/v d’un régu- de n-heneicosane est de 8 à 20 g par litre de com- lateur de croissance d’insectes dans de l’acétone. position.

12. Une composition "lure and kill" ("attirer et tuer") pour 21. Un procédé tel que revendiqué dans la revendication attirer des moustiques Aedes aegypti de manière à 25 20, dans lequel ladite concentration de n-heneico- ce qu’ils pondent leurs oeufs dans un site cible et sane est de 10 à 12 g par litre de composition. pour tuer les larves de moustiques ou réguler la croissance des larves de moustiques, qui comprend 22. Un procédé tel que revendiqué dans l’une des revendications 17 à 21, dans lequel à l’étape (a) le n- a) une quantité efficace de n-heneicosane, et 30 heneicosane est dissous dans de l’éther de pétrole, b) une quantité efficace d’un régulateur de crois- du n-pentane, du n-hexane, du méthanol ou du di- sance d’insectes, chlorométhane. dans un support inerte. 23. Un procédé tel que revendiqué dans la revendication 13. Une composition selon la revendication 12, dans la- 35 20, qui comprend le fait de : quelle le régulateur de croissance d’insectes est sé- lectionné parmi le diflubenzuron, le neemazal et le (a) préparer une solution attractive pour la ponte triflumuron. des oeufs en dissolvant de 8 à 20 g de n-heneicosane dans 500 ml de solvant organique, 14. Une composition selon la revendication 12 ou la re- 40 (b) préparer une solution de régulation d’insec- vendication 13, qui est sous la forme d’un spray. tes en dissolvant de 800 à 2000 mg de régula- teur de croissance d’insectes dans 500 ml de 15. Une composition "lure and kill" ("attirer et tuer") pour solvant organique, et attirer des moustiques Aedes aegypti de manière à (c) mélanger intimement les deux solutions ci- ce qu’ils pondent leurs oeufs dans un site cible et 45 dessus. pour tuer les larves de moustiques ou réguler la croissance des larves de moustiques, qui comprend 24. Un procédé tel que revendiqué dans l’une des re- une quantité efficace de n-heneicosane dissoute vendications 17 à 19, qui comprend le mélange, dans un solvant organique approprié, et une quantité dans un rapport de 1:1, de 1,0 à 2% p/v de n-henei- efficace d’un insecticide. 50 cosane dans du n-pentane, du n-hexane avec de 0,1 à 0,2% p/v d’un régulateur de croissance d’in- 16. Une composition selon la revendication 15, dans la- sectes dans de l’acétone. quelle l’insecticide est un larvicide. 25. Une méthode pour le contrôle des moustiques Ae- 17. Un procédé pour la préparation d’une composition 55 des aegypti, qui comprend le fait d’attirer des mous- pour le contrôle du moustique transmetteur de la tiques de manière à ce qu’ils pondent des oeufs dans dengue Aedes aegypti, comprenant des étapes con- un site cible et de tuer les larves qui en résultent ou sistant à de réguler la croissance des larves de moustiques

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en traitant le site avec (a) une quantité efficace de n-heneicosane organique dissoute dans un solvant, et (b) (i) une quantité efficace d’un régulateur de croissance d’insectes dissoute dans un solvant organique, ou (ii)une quantité efficace d’un insecticide. 5

26. Une méthode selon la revendication 25, dans laquelle le traitement est effectué avec une composition selon l’une quelconque des revendications 2 à 11. 10 27. Une méthode selon la revendication 25 ou la revendication 26, dans laquelle les composants (a) et (bi) sont mélangés à l’avance ou juste avant l’utilisation.

28. Une méthode pour le contrôle des moustiques Ae- 15 des aegypti, qui comprend le fait d’attirer les moustiques de manière à ce qu’ils pondent des oeufs dans un site cible et de tuer les larves qui en résultent ou de réguler la croissance des larves de moustiques en traitant le site avec une composition telle que re- 20 vendiquée dans l’une des revendications 12 à 16.