Compositions and Methods for Controlling Insects

a technology of insects and compositions, applied in the field of insects control, can solve the problems of inability to identify non-target species, use of toxic or deleterious effects on mammals, fish, fowl or other species, and difficulty in identification, and achieve the effect of repelling insects

Inactive Publication Date: 2008-03-27
TYRATECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0112] Adult insect are randomly selected for testing the repellent effect of compositions and are not individually marked. About 5 insects per replicate are used. About 3 replicates are used for each treatment. Untreated control tests are included with only solvent (acetone) application to an equal sized population / replications, held under identical conditions. A filter paper (about 80 cm2) is treated with the composition (about 100 mg in 300 ml acetone). After about 3 min of air drying, the filter paper is placed in a dish and repellency against insects is performed. Insects are released to the dish, one insect at a time at the far end of the dish. Using one or more stopwatches, the time spent on either the filter paper or the untreated surface of the dish is recorded up to about 300 seconds. Repellency ratio (RR) is calculated as follows: RR=[(time on control surface−time on treated surface) / total time of test]. If RR>0 then the composition is considered to have a repellant effect, that is to say, an effect, wherein more insects are repelled away from treated surface than the control surface; if RR<0 then the composition is considered not to have a repellant effect.

Problems solved by technology

Most of the previously known and commercialized products having sufficient pesticidal activity to be useful also have toxic or deleterious effects on mammals, fish, fowl or other species which are not the target of the product.
Other compounds, which may be less toxic to mammals and other non-target species, are sometimes difficult to identify.

Method used

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  • Compositions and Methods for Controlling Insects
  • Compositions and Methods for Controlling Insects
  • Compositions and Methods for Controlling Insects

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Stably Transfected Schneider Cell Lines with Tyramine Receptor (TyrR)

[0057] A. PCR Amplification and Subcloning Drosophila melanogaster Tyramine Receptor

[0058] Tyramine receptor is amplified from Drosophila melanogaster head cDNA phage library GH that is obtained through the Berkeley Drosophila Genome Project (Baumann, A., 1999, Drosophila melanogaster mRNA for octopamine receptor, splice variant 1B NCBI direct submission, Accession AJ007617). The nucleic acid sequence and the peptide sequence of TyrR are set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively. Phage DNA is purified from this library using a liquid culture lysate. (Baxter, et al., 1999, Insect Biochem Mol Biol 29, 461-467). Briefly, oligonucleotides that are used to amplify the open reading frame of the Drosophila tyramine receptor (TyrR) (Han, et al., 1998, J Neurosci 18, 3650-3658; von Nickisch-Rosenegk, et al., 1996. Insect Biochem Mol Biol 26, 817-827) consist of the 5′ oligonucleotide: 5′gccga...

example 2

Treatment of Cells Expressing the Tyramine Receptor and Effect of Compositions on Intracellular [cAMP]

[0073] Cells are grown on dishes and the media changed the day before the treatment. When cells are approximately 95% confluent, media is aspirated and the cells are washed one time with about 5 mL of about 27° C. insect saline (170 mM NaCl, 6.0 mM KCl, 2.0 mM NaHCO3, 17.0 mM glucose, 6.0 mM NaH2PO4, 2.0 mM CaCl2, 4.0 mM MgCl2; pH 7.0). About 20 mL of insect saline is added, and cells are harvested by gentle scraping. An aliquot of the cells is counted by hemocytometer, and the cells are then centrifuged for about 5 minutes at about 1000 RPM. Cells are resuspended to give about 3×106 cells per mL. IBMX is added to about 200 μM. Then about 1 mL of cell suspension is aliquoted for treatment. Forskolin (cAMP inducing agent), tyramine or different composition candidates are added, and the cells are incubated at about 27° C. for about 10 minutes.

[0074] Treated cells are centrifuged at a...

example 3

Treatment of Cells Expressing the Tyramine Receptor and Effect of Compositions on Intracellular [Ca2+]

[0076] Intracellular calcium ion concentrations ([Ca2+]i) are measured by using the acetoxymethyl (AM) ester of the fluorescent indicator fura-2 (Enan, et al., Biochem. Pharmacol vol 51, 447-454). In this study, cells expressing tyramine receptor are grown under standard conditions. A cell suspension is prepared in assay buffer (140 mM NaCL, 10 mM HEPES, 10 mM glucose, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2) and cell number adjusted to about 2×106 cells per ml. Briefly, about 1.0 ml cell suspension (about 2×106 cells) is incubated with about 5 μM Fura 2 / AM for about 30 min at about 28° C. After incubation, the cells are pelleted at about 3700 rpm for about 10 sec at room temperature and then resuspended in about 1.5 ml assay buffer. [Ca2+]i changes are analyzed in spectrofluorometer in the presence and absence of test chemicals. Excitation wave lengths are about 340 nm (generated by Ca2+-...

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Abstract

The present invention comprises compositions methods and cell lines related to controlling insects. An embodiment of a composition comprises a plant essential oil and targets at least one receptor of insects chosen from tyramine receptor, Or83b olfactory receptor, and Or43a olfactory receptor, resulting in a change in the intracellular levels of cAMP, Ca2+, or both.

Description

1. CROSS REFERENCES TO RELATED APPLICATION [0001] This application is a divisional of commonly assigned and co-pending U.S. patent application Ser. No. 10 / 832,022 filed Apr. 26, 2004, which claims priority from U.S. Provisional Application Ser. No. 60 / 465,320 filed Apr. 24, 2003 and U.S. Provisional Application Ser. No. 60 / 532,503 filed Dec. 24, 2003. The entire disclosures contained in U.S. application Ser. No. 10 / 832,022 and U.S. Provisional Application Ser. Nos. 60 / 465,320 and 60 / 532,503 are incorporated herein by this reference.2. FIELD OF THE INVENTION [0002] The present invention relates to compositions, methods, cell lines and reports related to controlling insects. 3. BACKGROUND OF THE INVENTION [0003] Animals have chemosensory and mechanosensory systems that recognize a large array of environmental stimuli, generating behavioral responses. Behavioral studies have been conducted to understand the genetics of these systems. The olfactory system plays a role in the survival an...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01N65/00A01N27/00A01N31/02A01P7/04A01N37/00A01N37/18A01N31/04A01N31/08A01N31/16A01N35/02A01N35/06A01N37/44A01N43/30A01N49/00G01N33/566
CPCA01N27/00A01N31/04G01N2333/43552G01N33/566G01N33/5085A01N65/00A01N49/00A01N43/30A01N31/08A01N31/16A01N35/02A01N35/06A01N37/18A01N37/44A01N2300/00A01N25/02A01N53/00A01N65/08A01N65/10A01N65/22A01N65/44Y02A50/30A01N65/32A01N65/36
Inventor ENAN, ESSAM
Owner TYRATECH
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