Patent Application: US-95237707-A

Abstract:
this invention relates to novel apparatus and methods which use specific wavelengths of visible light , or combinations of specific wavelengths of visible light with specific wavelengths of ultra - violet light , to manipulate the behavior of stored product insect pests , including moths and indian meal moths . the apparatus for attracting stored product insect pests , including the indian meal moth , plodia interpunctella , the mediterranean flour moth , ephestia kuhniella , the tobacco moth , ephestia elutella , the almond moth , cadra cautella , and the raisin moth , cadra figulielella , consists of a light source placed in a trap .

Description:
throughout the following description specific details are set forth in order to provide a more thorough understanding to persons skilled in the art . however , well known elements may not have been shown or described in detail to avoid unnecessarily obscuring the disclosure . accordingly , the description and drawings are to be regarded in an illustrative , rather than a restrictive , sense . indian meal moth ( imm ) larvae , plodia interpunctella , were obtained from infested cereal bars provided by pherotech international inc . larvae were reared at 27 ° c . at a photoperiod of 17 hours light and 7 hours dark . the rearing medium was modified from le cato ( 1976 ) and consisted of whole wheat flour ( 27 . 5 % by volume ), yellow cornmeal ( 27 . 5 %), purina one dog food ( 13 . 5 %), brewers yeast ( 6 . 9 %), honey ( 6 . 9 %), glycerine ( 6 . 9 %; 96 % pure ), quaker rolled oats ( 6 . 8 %) and wheat germ ( 3 . 4 %). fifth instar larvae were separated by sex and placed in groups of 12 - 15 specimens into petri dishes ( 10 cm diam ), containing corrugated cardboard as pupation sites . eclosed adults were kept under the same conditions as larvae ( see above ). to obtain gravid females , 3 - 4 virgin males and 2 - 3 virgin females were kept together during the dark phase in small cages ( 10 × 10 × 10 cm ). the next day , females were assumed mated and were used for colony rearing or laboratory experiments . all adult moths used in laboratory experiments were 2 - 5 days old . still - air , two - or four - choice laboratory bioassays ( experiments 1 - 10 ) were conducted in a modified wind tunnel ( 1 × 1 × 3 m long ) with air entry and exit sections covered by mesh screens , and also by black paper in experiments 4 - 10 to minimize light reflection ( fig1 ). fig1 illustrates the scheme of the experimental design employed in two - choice or four - choice experiments . traps , the platform for releasing moths and light - emitting - diodes ( led ) are drawn not to scale . for each replicate , two petri dishes with ≦ 5 insects each were placed on a 50 - cm tall , black felt - covered platform ( 23 × 30 cm ) in the centre of the tunnel . in two - choice experiments , a green delta trap ( pherotech international inc .) was affixed to a metal pole at a height of 50 cm , and was positioned to the left and right of , and 1 . 5 m apart from , the release platform . in four - choice experiment 7 , the design was identical except that one trap was near ( 30 cm ; 45 ° angle ) each corner of the tunnel . light sources as test stimuli were randomly assigned to , and mounted within , traps . all experiments were conducted in the first 2 hours of the 7 - hour dark phase , when imms forage for mates or suitable oviposition sites . an experimental replicate was initiated by lifting the lid of each petri dish on the release platform , and was terminated by scoring the number of moths captured in each trap two hours later . all moths not responding were removed from the wind tunnel prior to initiating a new experimental replicate . after each set of three replicates , the wind tunnel was wiped with 70 % ethanol and left to “ aerate ” overnight . relative attractiveness of white , red , green and blue light sources to mated females tested in two - choice experiments to determine the spectrum of visible light that is most effective in attracting imms , a 6 - volt light bulb connected to a 9 - volt power source was placed inside a 2 - ml glass vial ( 10 × 28 mm ) which filtered out ultraviolet light . the glass vial , in turn , was surrounded by a cylindrical ( 5 . 5 × 10 . 5 cm diam ) flexible filter ( lee filters , hamshire , england ) that generated light spectra in the blue range ( 400 - 475 nm , peaking at 400 , 425 and 450 nm ; referred to as “ rose purple 7 ”), green range ( 475 - 600 nm , peaking at 510 nm , 545 nm , and 575 nm ; “ lime 8 ”) and red range ( 575 - 750 , peaking at 610 nm and 655 nm ; “ orange 9 ”). a clear filter ( heat shield # 269 ) was used to generate white light . light intensities were measured with a mastersix photometer ( gossen foto - und lichtmesstechnik , nürnberg , germany ) with the diffuser removed so that low - light levels could be measured . all light sources were tested at an intensity of 53 - 170 lux . in two - choice experiments 1 - 3 , it was found that gravid female imms preferred blue light over red light ( experiment 1 ), and white light ( containing blue light ) over red light ( experiment 2 ), but failed to show a significant preference for red or green light , which were equally unattractive ( experiment 3 ) ( see fig2 ). this unexpected result shows for the first time a preference for visible light of a defined wavelength over white light that includes that defined wavelength . sambaraju and phillips ( 2006 ) showed this for ultraviolet , but not visible , light . fig2 illustrates graphical data of mated female indian meal moths responding in still - air , two - choice laboratory experiments 1 - 3 to various light sources , each tested at a light intensity of 53 - 170 lux . in each experiment , an asterisk (*) indicates a statistically significant preference for the respective test stimulus ; wilcoxon paired - sample test , p & lt ; 0 . 05 . relative attractiveness of various wavelength ranges to males , virgin females and mated females tested in four - choice experiments to further determine the spectrum of visible light that is most effective in attracting imms , four - choice experiments were conducted . modified desk lamps ( espressivo , ikea ) with 20 - watt halogen bulbs were used as light sources to test the response of males ( experiment 4 ), virgin females ( experiment 5 ) and mated females ( experiment 6 ). each desk lamp was connected to a rheostat to adjust light intensity , and the halogen bulb was fitted with a black cardboard cylinder ( 8 × 12 cm wide ), with the light filter mounted at the front 8 cm apart from the bulb . the cylinder projected the light in one direction . flexible filters ( lee filters , hamshire , england ) that generated light spectra in the blue range ( 400 - 475 nm , referred to as “ rose purple 7 ”), green range ( 475 - 600 nm , “ lime 8 ”), orange range ( 525 - 750 nm , “ orange 9 ”) or red range ( 590 - 800 nm , “ light red ”). filter spectra and light intensities were measured with an hr4000 high - resolution spectrometer ( ocean optics dunedin fla .). all light sources were tested at an intensity of 15 μw / cm 2 integrated from 350 - 700 nm measured at the filter , 8 cm from the halogen light source . in four - choice experiments 4 - 6 , it was found that males and mated females showed a significant preference for blue light over red , green and orange light , but that virgin females had no preference for any wavelength range ( fig3 ). fig3 illustrates graphical data of male , mated female and virgin female indian meal moths responding in still air , four - choice laboratory experiments 4 - 6 to various light sources , each emitting 15 μw per 1 cm 2 . in each experiment , bars with different letters are significantly different ; analysis of variance with tukey &# 39 ; s test for multiple comparison of means , p & lt ; 0 . 05 . to determine the intensity of blue light ( 400 - 475 nm ) most effective in attracting gravid female imms , light intensities of 50 versus 200 lux ( experiment 7 ), 200 versus 1 , 000 lux ( experiment 8 ), and 1 , 000 versus 3 , 000 lux ( experiment 9 ) were tested in two - choice experiments . in all three experiments , the filter “ rose purple 7 ” ( see above ) was used to generate blue light , but the light sources differed . in experiment 7 , the light source consisted of a 6 . 4 - volt bulb connected to a 100 - ohm adjustable resistor powered at 9 volts . experiments 8 and 9 deployed a modified desk lamp ( espressivo , ikea ) with a 20 - watt halogen bulb to generate light intensities of 1 , 000 lux and 3 , 000 lux . the desk lamp was connected to a rheostat to adjust light intensities . the halogen bulb was fitted with a black cardboard cylinder ( 8 × 12 cm wide ), with the filter “ rose purple 7 ” mounted at the front , 8 cm apart from the bulb . the cylinder projected the light in one direction . in each of experiments 7 - 9 , it was found that mated female imms preferred the blue light of greater intensity over that of lower intensity ( fig4 ). fig4 illustrates graphical data of mated female indian meal moths responding in still - air , two - choice laboratory experiments 7 - 9 to blue light ( 400 - 475 nm ) of different intensities . in each experiment , an asterisk (*) indicates a statistically significant preference for the respective test stimulus ; wilcoxon paired - sample test , p & lt ; 0 . 05 . attractiveness of specific wavelengths in the blue light spectrum ( 400 - 475 nm ) each tested at a light intensity of 200 lux to determine the wavelength in the blue light spectrum ( 400 - 475 nm ) most effective in attracting mated female imms , light - emitting - diodes ( led ; roithner lasertechnik , vienna , austria ) with peak wavelengths of 405 nm ( range : 400 - 410 nm ), 435 nm ( range : 410 - 470 nm ), 450 nm ( range : 440 - 460 nm ) and 470 nm ( range : 465 - 475 nm ) were tested in four - choice experiment 10 . for each replicate , one of the four leds was randomly assigned to , and mounted within , one of four green delta traps ( see general experimental design ; fig1 ), using a resistor to adjust the intensity of each led to 200 lux . in experiment 10 , it was found that the led with peak wavelength 405 nm was significantly more effective than leds with peak wavelength 435 nm , 450 nm or 470 nm in attracting gravid female imms . the latter three peak wavelengths were equally unattractive to female moths ( see fig5 ). fig5 illustrates graphical data of mated female indian meal moths responding in still - air , four - choice laboratory experiment 10 to specific wavelengths ( 405 , 435 , 450 or 470 nm ) in the blue - light wavelength range ( 400 - 475 nm ). bars with different letters are significantly different ; analysis of variance with tukey &# 39 ; s test for multiple comparison of means , p & lt ; 0 . 05 . attractiveness of specific wavelengths in the blue - light spectrum ( 400 - 475 nm ) each tested at a light intensity of 200 μw per 1 cm 2 to further determine the wavelength in the blue light spectrum most effective in attracting mated females , an additional four - choice experiment ( experiment 11 ) was conducted . the experimental design was identical to that of experiment 10 except that the leds were calibrated to emit 200 μw per 1 cm 2 , integrated from 350 - 550 nm using a hr4000 high - resolution spectrometer ( ocean optics dunedin fla .). in experiment 11 , it was found that the led with peak wavelength 405 nm was significantly more effective in attracting gravid females than were leds with peak wavelength 435 nm , 450 nm or 470 nm . the latter three peak wavelengths were equally unattractive to female moths ( fig6 ). fig6 illustrates graphical data of mated female indian meal moths responding in still - air , four - choice laboratory experiment 11 to specific wavelengths ( 405 , 435 , 450 or 470 nm ) in the blue - light wavelength range ( 400 - 475 nm ), each calibrated at 200 μw per 1 cm 2 . bars with different letters are significantly different ; analysis of variance with tukey &# 39 ; s test for multiple comparison of means , p & lt ; 0 . 05 ; led = light emitting diode . attractiveness of “ led 405 ” and the blue - light spectrum 400 - 475 nm to males , virgin females and mated females to compare the relative attractiveness of blue light ( 400 - 475 nm ) and specific wavelength 405 nm (+/− 5 nm ), both light sources at 200 lux each were tested in two - choice experiments 8 - 10 , with males ( experiment 12 ), virgin females ( experiment 13 ) and mated females ( experiment 14 ) as bioassay insects . the blue - light spectrum was generated from a desk lamp ( espressivio , ikea ) with a halogen bulb , fitted with a cardboard cylinder ( 8 × 12 . 5 cm diam ) for projecting the light and carrying the filter “ rose purple 7 ” ( as described for experiments 4 - 6 ). to standardize visual stimuli , the same set - up was used for the “ 405 - nm led ” which was mounted just in front of the turned - off halogen bulb . in two - choice experiments 12 - 14 , it was found that males ( experiment 12 ), virgin females ( experiment 13 ) and mated females ( experiment 14 ), all preferred the led with peak wavelength 405 nm over the blue - light spectrum 400 - 475 nm ( fig7 ). this result was surprising and unexpected , given that indian meal moths , and other stored product moths , were not highly attracted to light of an almost identical wavelength ( 404 . 7 nm ), which was described by stermer ( 1959 ) as an “ unattractive waveband ”. fig7 illustrates graphical data of male , virgin female and mated female indian meal moths responding in still - air , two - choice laboratory experiments 12 - 14 to a source of blue light ( 400 - 475 nm ) and a specific wavelength ( 405 nm ) each tested at an intensity of 200 lux . in each experiment , an asterisk (*) indicates a significant preference for the respective test stimulus ; wilcoxin paired - sample test , p & lt ; 0 . 05 . attractiveness of specific wavelengths in the green - light spectrum ( 505 - 572 nm ) each tested at a light intensity of 30 μw per 1 cm 2 considering that green light ( 475 - 600 nm ) was somewhat attractive ( although not statistically significant ) to males , virgin females and mated females in experiments 4 - 6 ( see fig3 ), and that attraction of imm to green light is reported in the literature ( stermer 1959 ; soderstrom 1970 ; kirkpatric and marzke 1970 ), four - choice bioassays ( experiments 15 - 17 ) were designed to determine the specific wavelength ( s ) responsible for the attractiveness . light - emitting - diodes ( led ) with peak wavelengths of 505 nm , 525 nm , 565 nm or 572 nm ( roithner lasertechnik , vienna , austria ) were deployed to test the response of males ( experiment 15 ), virgin females ( experiment 16 ) and mated females ( experiment 17 ). for each replicate , one of the four leds was randomly assigned to , and mounted within , one of four green delta traps ( see general experimental design ; fig1 ), adjusting with a resistor the intensity of each led to 30 μw per 1 cm 2 . in experiments 15 - 17 , it was found that there was a weak preference by males , virgin females and mated females to leds emitting a peak wavelength of 505 nm or 525 nm ( fig8 ). in experiment 15 , the 525 - nm led attracted significantly more males than did the 572 - nm led . in experiment 17 , the 525 - nm led attracted significantly more mated females than did the 565 - nm led ( fig8 ). fig8 illustrates graphical data of male , virgin female and mated female indian meal moths responding in still - air , four - choice laboratory experiments 15 - 17 to light emitting diodes ( leds ) emitting at 30 μw per 1 cm 2 a peak wavelength of 505 , 525 , 565 or 572 nm . bars with different letters are significantly different ; analysis of variance with tukey &# 39 ; s test for multiple comparison of means , p & lt ; 0 . 05 . attractiveness of wavelength 405 nm (+/− 5 nm ) tested alone or in combination with wavelength 350 nm (+ 5 /− nm ) to determine whether attraction of mated female imms to the wavelength 405 nm would increase in the presence of another specific wavelength , two - choice experiment 18 ( see fig1 ) tested a single led emitting peak wavelength 405 nm at 200 μw per 1 cm 2 versus two leds , one of which emitting the peak wavelength 405 nm at 180 μw per 1 cm 2 and the other led emitting the peak wavelength 350 nm at 20 μw per 1 cm 2 . great care was taken to adjust to 200 μw per 1 cm 2 the total light intensity emitted from either the single led or the paired leds . for each replicate , the paired leds were positioned on top of each other and mounted within a green delta trap , using a resistor to adjust the intensity of each led . the position of test stimuli ( see fig1 ) was alternated between replicates . in experiment 18 , it was found that the paired 405 - nm and 350 - nm leds attracted more mated females than did the single 405 - nm led ( fig9 ). this indicates that attraction of imms to a 405 - nm led can be improved by addition of other specific wavelengths . fig9 illustrates graphical data of mated female indian meal moths responding in still - air , two - choice laboratory experiment 18 to a single wavelength ( 405 - nm led ) or to a wavelength combination ( paired 405 - nm led and 350 - nm led ), with single or combined wavelength stimuli tested at identical light intensity ( 200 μw per 1 cm 2 ). in all experiments which tested the effective wavelength 405 ± 5 nm , a large proportion of all moths released into the modified wind tunnel ( see fig1 ) were captured within just two hours , indicating that this technology has great potential for suppression of imm populations in private households and industrial settings . while a number of exemplary aspects and embodiments have been discussed above , those of skill in the art will recognize certain modifications , permutations , additions and sub - combinations thereof . it is therefore intended that the following appended claims and claims hereafter introduced are interpreted to include all such modifications , permutations , additions and sub - combinations as are within their true spirit and scope . barrer , p . m . 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