PESTICIDE COMPOSITION AND METHOD FOR CONTROLLING THE MEDITERRANEAN FRUIT FLY AND THE ORIENTAL FRUIT FLY

Abstract: A composition and method for eradicating or suppressing a population of Mediterranean fruit flies or oriental fruit flies which introduces substantially minimal safety risks to humans, agriculture and nontargeted living creatures includes: providing an insecticide composition which includes at least one photoactive dye which is preferably a mixture of Phloxine B and uranine; causing the composition to be ingested by the targeted fruit fly population by spraying or providing the insecticide at a location where the fruit flies feed; and whereby ingestion of the photoactive dye component of the composition by a Mediterranean fruit fly or oriental fruit fly causes a toxic quantity of photons to penetrate within the body of the fly.

Description: PESTICIDE COMPOSITION AND METHOD FOR CONTROLLING THE MEDITERRANEAN FRUIT FLY AND THE ORIENTAL FRUIT FLY This is a continuation-in-part of application Serial No. 08/353,726 filed December 12, 1994. Background of the Invention The fruit flies of the family Tephritidae include several species tha are major pests of agriculture throughout the world and that represent serious threat to U.S. agriculture. The U.S. Department of Agricultur (USDA), Animal and Plant Health Inspection Service (APHIS), i cooperation with other Federal and State organizations, has conducted number of programs to eradicate some species of fruit flies when thes insects have been introduced. These programs generally have employe an integrated pest management approach to eradication. Many recen programs have involved application of malathion bait spray to effectivel lower fly populations in the infested area followed by release of steril flies. Aerial applications of the bait spray over populated areas to contr infestations of fruit flies have been controversial. Concerns about advers health effects from exposure to malathion bait spray have been raised b residents of treated neighborhoods. The currently used malathion bai system for control of Mediterranean fruit flies contains as the main ingredients, Nu-Lure (acid hydrolyzed protein) as an attractant food bait and the contact insecticide, malathion. This system has a bad public perception, damages paint finishes on cars, and, because of the high concentration of insecticides (10-20%) in the bait to ensure temporal stability, this insecticide is extremely detrimental to beneficial insects that may contact the bait surface or be exposed to volatile fumes after application. The United States government has mandated that certain currently listed pesticides, such as malathion, should have a more restricted use pattern. The Environmental Protection Agency has specifically requested that safer pesticides be developed for use in the agricultural sector. Summary of the Invention In the quest for these aforementioned safer pesticides, it has been unexpectedly discovered that dye-sensitized photoactive substances can be effectively utilized as the active ingredient in insecticides for use in a bait station or as bait spray component which target Mediterranean fruit flies and many other tephritid fruit flies. The preferred photoactive dye for these purposes is a halogenated xanthene such as phloxine B or a mixture of a halogenated xanthene and uranine. Other known xanthen dyes which could be used include erythrosin B and rose bengal. Thes dyes are more fully described in the U.S. patents to Crounse and Heitz U.S. Patent No. 4,647,578 and U.S. Patent No. 4,320,140, the subjec matter of which is incorporated herein by reference. When employin halogenated xanthene dyes, light activated toxicity is especially marke with respect to Mediterranean fruit flies and oriental fruit flies. Th photoactive dye ingredient of the present invention is effective due to th release of light induced free radical molecules within the body of th insect which has ingested the composition containing the dye. Due to it small proportionate body mass, the dose of light is fatal to the insect. Although being extremely toxic with respect to the targete Mediterranean fruit fly or oriental fruit fly, the preferred dyes of th instant invention are non-toxic to humans, most mammals, an nontargeted insects. This is a substantial advance in the industry, a prior art pesticides which were adapted for the purpose of controlling frui fly populations, namely malathion, have posed health risks to humans Because malathion is highly penetrating and invades cells very quickl upon contact, malathion is highly potent as a poison, both to insects an nontargeted species. The present invention, on the other hand, i directed to an active ingredient which is toxic to the targeted Mediterranean or oriental fruit fly, not due to cell penetration, but rather due to exposure to a quantity of light which is fatal to the fly. The effect of the same quantity of exposure by a human or other mammal would be negligible. In this regard, the compound phloxine B (2',4',5',7'-tetrabromo-4,5,6,7-tetrachlorofluorescein) has been consumed by humans for decades. It is registered as D & C Red No. 28 as a drug and cosmetic additive and has been included into such commercially available products as Pepto Bismol. In fact, the intrinsic toxicity of malathion is 62.5 times greater than that of phloxine B as measured by acute LD 50 in rats. Additionally, unlike malathion which kills any insect merely upon contact, even nontargeted species such as honeybees, the photoactive dye ingredient of the present invention is not harmful unless ingested. Indeed, the skin penetrability of phloxine B has been calculated to be about 87 times lower than that of malathion by octanol/water partition coefficient analysis conducted at the USDA. Furthermore, because the active xanthene dye component is light-activated, a much smaller quantity is necessary to provide the desired effect. Moreover, contrary to the prior art broad spectrum pesticides which have a half-life on the order of months or even years the xanthene based ingredient will naturally photodegrade in th environment within hours or, at the most, days. Thus, not only provid an extremely high kill rate when introduced in the targeted Mediterranea or oriental fruit fly populations, but additionally pose little or no risk fo use in the agriculture industry. An additional benefit which is offered by the use of xanthene base dyes as the active ingredient in pesticide compositions, resides in the fac that, after ingestion, the dye does not kill the insect immediately. Thi allows the insect which has ingested the pesticide to effectively transfe some quantity of the pesticide among other insects in the population Therefore, substantially more insects will be eliminated due to the transfe of the toxic material between flies. Another aspect of the invention is use of phloxine B in combinatio with uranine. Uranine is registered as D & C Yellow Dye No. 8 for use a a color additive in drugs and cosmetics. The present insecticidal composition further contemplates th inclusion of an attractant bait which has the following properties: i attracts the target fruit flies to the location of the bait, and it stimulate the target fruit flies to feed on the bait. Attractant baits include (1 ) carbohydrate sugar source, for example, sugars such as sucrose or fructose, or sugar substrates such as molasses or honey; (2) a proteinaceous food bait such as hydrolyzed protein (either by acid hydrolysis or enzymatic action), or (3) a combination of carbohydrate sugar source and hydrolyzed protein bait. The composition may optionally include anti-foaming agents and other adjuvants to increase the permeability of the ingested dye within the gut of the insect. It is therefore an important object of the present invention to provide a pesticide that is toxic only when ingested by particular species of insects and in particular the Mediterranean or oriental fruit fly and other tephritid fruit flies. It is a further object to provide a pesticide that has a delayed toxicity to the insect. A still further object lies in the provisions of a pesticide which has minimal detrimental effect on the behavior of the insect between the period of ingestion and the onset of toxicity. A still further object of the present invention lies in the provision of an insecticidal composition which is highly attractive to both sexes of the Mediterranean and oriental fruit fly. A yet further object of the present invention resides in the provision of an insecticidal composition which contains no feeding inhibitors th may limit feeding or may induce a learned behavior to avoid the bait. Lastly, it is an object of the present invention to provide a insecticidal composition which incorporates a bait that stimulates feedin and induces engorgement by the insects. These together with other objects of the invention, are pointed o with particularity in the following detailed description and claims annexe hereto and forming part of this disclosure. Brief Description of the Drawings Figure 1 shows percent mortality of adult Mediterranean fruit flie as a function of varying concentrations of 1 :1 molar mixture of phloxin B and uranine and time after feeding. Figure 2 shows the functional relationship between concentratio of dyes in 10% molasses and mortality of Mediterranean fruit fly adult Figure 3 shows the functional relationship between concentratio of dyes in 1 % Nu-Lure and mortality of Mediterranean fruit fly adults. Figure 4 shows the functional relationship between concentratio (M) of 1 :1 molar mixture of phloxine B and uranine in 1 % Nu-Lure an mortality of oriental fruit fly adults. Figures 5A and 5B show percent mortality of adult oriental fruit fly males and females, respectively, as a function of varying concentrations of 1 :1 molar mixture of phloxine B and uranine and time after feeding. Figure 6 shows percent mortality of wild oriental fruit fly adults as a function of methyl eugenol alone, methyl eugenol plus a 1 :1 molar mixture of phloxine B and uranine, and methyl eugenol plus naled, and time after exposure to light. Figure 7 shows the increase in mortality over time of wild oriental fruit fly adults exposed to methyl eugenol alone and methyl eugenol plus a 1 :1 molar mixture of phloxine B and uranine at two concentrations. Figure 8 shows the increase in mortality over time of wild oriental fruit fly adults exposed to methyl eugenol alone and methyl eugenol plus phloxine B at two concentrations. Detailed Description of the Invention Toxicity of several substituted xanthene dyes to many species o insects has been documented in the laboratory and field (Heitz, Chapte 1 , Lioht Activated Pesticides. J. R. Heitz and K. R. Downum (eds.), American Chemical Society Symposium Series 339, American Chemica Society, Washington, D.C., 1987). Among these are the dye-sensitize photoactive toxicity of phloxine B to adult housefly, Musca domestica L (Fondren and Heitz, Environmental Entomology 8:432-436, 1979), fac fly, M. autumnalis De Geer (Fondren and Heitz, Environmenta Entomology 7:843-846, 1978), boll weevil, Anthonomus orandi Boheman (Callaham et al., Environmental Entomology 4:837-841 , 1975) black cutworm, Aorotis ipsilon (Clement et al., Journal of Economi Entomology 73:390-392, 1980), and imported fire ant, Solenoosis richter (Forel) (Callaham et al., Comp. Biochem. Phvsiol. 51 :123-128, 1975 Broome et al., Environmental Entomology 4:883-886, 1975). Ligh activated toxicity has also been reported in uranine-fed adult house fl (Fondren and Heitz, 1979, supra), face fly (Fondren and Heitz, 1978 supra), and imported fire ant (Callaham et al., 1975, supra). Althoug dye-sensitized photooxidative toxicity from uranine is considerably les than phloxine B, uraπ' e has been found to cause synergistic effect when combined with other xanthene dyes. Phloxine B (2',4',5',7'-tetrabromo-4,5,6,7-tetrachlorofluorescein) is registered for use as a color additive in drugs and cosmetics. Uranine, chemically known as fluorescein 3',6'-dihydroxyspiro[isobenzofuran-1 (3H,9'- [9H]xanthen]-3-one, disodium salt, is registered for use as a color additive in cosmetics as D & C Yellow No. 8. The toxicity of phloxine B combined with uranine is of importance in finding suitable alternatives to insecticides that are mixed with food baits in sprays that are used in eradication or suppressing many tephritid fruit fly populations. Thus, in an effort to find an environmentally acceptable alternative to malathion, we determined the toxicity of a mixture of phloxine B and uranine to Mediterranean fruit fly and oriental fruit fly adults. Data presented here are fundamental information in developing bait formulations, containing phloxine B and uranine for controlling Mediterranean fruit fly and other tephritid fruit fly pests. As discussed above, it is contemplated that the present insecticidal composition includes an attractant bait which attracts the target insect and stimulates it to feed on the bait. Attractant baits include 1 ) a carbohydrate sugar source, for example, sugars such as sucrose or fructose or complex sugar substrates such as molasses or honey; 2) a proteinaceous food bait such as hydrolyzed protein (either by aci hydrolysis or enzymatic action); 3) a combination of sugar source an hydrolyzed protein bait or 4) methyl eugenol. Examples of hydrolyzed protein sources are the commerciall available preparation, Nu-Lure™ (44% corn gluten meal, hydrolyzed, an 56% inert ingredients, Miller Chemical and Fertilizer, Hanover, PA Mazoferm™ (condensed fermented corn extractives, E802, Corn Product Summit-Argo, II.); hydrolyzed torula yeast, and hydrolyzed brewers yeas The dye and bait formulation can be used in dispensers set out i feeding bait stations or used in a sprayable formulation for ground or a applications. The formulation may be applied by various types equipment from hand held sprayers to high pressure ground sprayers t helicopters to fixed-wing aircraft. To accommodate for the different spray volume delivery rate varying amounts of water is used as a diluent to the active ingredients the spray formulation. An exemplary formulation is: photodye from 0.1 to 1 % of total formulation hydrolyzed protein from 35.0 to 99.0% of total formulatio sugar source from 0.0 to 20% of total formulation added water from 0.0 to 70% of total formulation O 96/18295 PCI7US95/ 15962 The total formulation should be applied so as to obtain good spray coverage with from 0.5 to 2.0 pounds of hydrolyzed protein per treated acre. Another exemplary formulation is: photodye from 0.1 to 1 % of total formulation sugar source from 5 to 20% in water A dye plus bait formulation using methyl eugenol as the attractant bait is suitable for control of oriental fruit flies in male annihilation eradication programs. Male annihilation treatments are most often applied as spot applications of paste-like globs (generally about 600 spot applications per square mile) or as solutions adsorbed into solid carriers such as cigarette filter tips or 5 cm square fiberboard blocks hung on trees at rates of 500 to several thousand per square mile. As known to those in the art, any specific methyl eugenol formulation and use pattern is chosen based on various factors including accessibility and/or whether it is applied in an inhabited area. An exemplary formulation is: photodye from 0.5 to 5.0% of total formulation by wt. methyl eugenol from 79.5 to 70.0% of total formulation Min-U-Gel 400 from 20 to 25 % of total formulation (attapugite clay) The total formulation is applied as about 10 to 1 5 gram globs on tree trunks, telephone poles, etc. at a rate of about 600 spots per squar mile about every two weeks. It can also be applied as a thickened spra by aircraft at a rate of about 20 lbs per square mile every 1 to 2 week Another exemplary formulation is: photodye from 0.5 to 10.0% of total formulation by wt. methyl eugenol from 99.5 to 90.0% of total formulation This liquid formulation is adsorbed into various solid carriers such a cotton strings or wicks, cigarette filter tips, or cellulosic fiber wallboar cut into pieces of various sizes. These saturated solid carrier pieces ca be distributed by aircraft or hung on foliage at a rate so that the tot amount of liquid formulation is from about 20 to 100 lbs per square mil Generally, applications are repeated every 2 to 8 weeks. This liquid formulation can also be sprayed without any thickener solid carrier at rates of 20 lbs per square mile in aerial applications a about weekly intervals. The following examples describe the various innovative aspects o the present invention. Materials and Methods Insects: Mediterranean fruit fly pupae and Oriental fruit fly pupae wer obtained from the mass rearing facility of the USDA-ARS, Honolulu Pupae were kept in an insectary at 75 + 5 degrees C, 60 - 75% relativ humidity, and a 12:1 2 h (L.D.) photoperiod. Test adults were held under th same temperature, relative humidity, and photoperiod as the pupae, an were fed with water (in agar, consisting of 9.975 part water and 0.025 part agar (Gelcarin™) and diet consisting of 3 parts sucrose, 1 par protein yeast hydrolysate and 0.5 part Torula yeast. Five-d old adult were used in the succeeding experiments. Example 1 : Determining Variation in Medite rranean Fruit Fly Adul Mortality due to Concentration of Dyes, Sex, and Time afte Feeding. Phloxine B and Uranine Mixed in 1 0% Molasses. Fiv dilutions (percentage by weight) consisting of 0.001 %, 0.01 %, 0.1 %, 1 .0 and 10.0% of 1 :1 molar mixture of phloxine B (829 g per mole) and uranin (376 g per mole) in a stock of 10% molasses were prepared. Test adul flies confined in feeding chambers were provided with dye-molasse preparations saturated in cotton wicks; control flies were provided wit 10% molasses. Each feeding chamber had either 20 males or 20 females Adult feeding with dye-molasses mixtures commenced at 06.00 h an terminated at 08:00 h, providing a 2-h feeding duration. From the onset o feeding, each feeding chamber was kept 10 cm below two high intensity cool fluorescent lights yielding surface intensity of 1 8,000 LUX; the sam lighting condition was maintained throughout the 1 2-h light phase of th experiment duration. Following removal of wicks containing the dyes flies were provided with water and a diet consisting of sucrose, protei hydrolysate, and torula yeast. Mortality counts were made every hour fo the first 4 hours following feeding, at 1 8:00 h when lights wer automatically turned off, and during each succeeding morning at 8:00 h fo 5 d. The experiment followed a randomized complete block design with replicates. Adults used in the experiments were provided with only wate 24 h (i.e., 1 2-h dark and 12-h lights periods) prior to their use in th experiment. Results Fig. 1 shows the Variation in Mediterranean fruit fly adul mortality due to concentration of dyes, sex and time afte feeding. Phloxine B and Uranine were mixed in 10% Molasses Mortality of Mediterranean fruit fly adults fed with variou concentrations (by weight) of 1:1 molar mixture of phloxine B and uranin differed significantly with dose, sex, and number of hours afte termination of feeding. Mortality of both males and females increase with the increase in concentration of dyes from 0.001% to 0.1%; however at 1% to 10% concentrations, adult mortality was lower than tha observed at 0.1%. Five days after feeding molasses with phloxine B an uranine, the trend in mortality by dose for males was 0.1% > 0.01% = 1% 10% > 0.001% = 0%; for females, the mortality trend was 0.1% > 0.01% 1% > 10% > 0.001% > 0%. The cumulative mortality of females wad highe than that of males for all concentrations, except at 10%. There was n significant increase in mortality 24 h after feeding on molasses wit dyes for females and after 48 hours for males. Example 2 Determining Functional Relationship Between Concentratio of Dyes and Mortality of Mediterranean Fruit Fly Adults Phloxine B and Uranium Mixed in 10% Molasses. Based on result obtained in study 1, concentration-mortality relationship was determine using the following molarity (M) concentrations of 1:1 molar mixture o phloxine B and uranine in 10% molasses: 2.5 x 10 "5 , 5 x 10 ' 5 , 1 10- 4 , 2 x 10 "4 , 4x 10 "4 , 8x l0- 4 , 1.6x l0 "3 , 3.2x l0 "3 , 6.4X 10 ' 3 and 1.28 x 10" 2 . The assay procedure was similar to the methodolog described above. Control flies were fed with 10% molasses. Based o results of study 1, mortality counts were done only up to 2 d. Twent groups of 20 flies (by sex) were tested for each dye concentration. Adult used in the test were not starved (i.e., 24 h prior to their use in th experiment, adults had sugar-protein-water diet during the 12 h ligh phase and were provided with water only during the (12 h dark phase). Results Figure 2 shows the functional relationship betwee concentration of dyes and mortality. Phloxine B and uranin were in 10% molasses. Mortality of both males and females increase with the increase in concentration of dyes from 2.5 x 1 0' 5 M to 1 .6 10 " 3 M; however, at 3.2 x 1 0 * 3 M to 1.28 x 1 0 " 2 M, adult mortality wa lower that that observed at 1 .6 x 1 0" 2 M. The LC 50 was 1 .7 x 1 0" 3 M fo males and 1.3 x 1 0 ' 3 M for females. Both sexes were deterred fro feeding at concentrations greater than 1 .6 x 10 " 3 M. The LT 50 at 1 .6 1 0 " 3 M was about 7 hours for both sexes. Example 3 Phloxine B and Uranine Mixed in 1 % NuLure. Various dilution of dyes were prepared in a stock of 1 % NuLure (a commercial hydrolyze protein bait preparation) 44% corn gluten meal, hydrolyzed and 56% iner ingredients, Miller. Concentration mortality relationship was determine using the following molarity (M) concentrations: 2.5 x 1 0 " 5 , 1 x 10 " 4 4 x 1 0" 4 , 8 x 1 0 " 4 , 1.6 x 1 0 " 3 . Control flies were fed with 1 % NuLure The assay procedure was similar to the method described above, excep that fourteen groups of 20 females were tested for each dy concentration. Test adults were not starved (i.e. 24 h prior to their use i the experiment, adults had sugar-protein-water diet during the 1 2 h ligh phase and were provided with water only during the 1 2 h dark phase). R es u lts Figure 3 shows that when the food carrier for dye was 1 % NuLur (i.e., with relatively much lower feeding stimulant than molasses), ther was no significant increase in mortality from 2.5 x 10" 5 to 4 x 10" significant increase in mortality was observed from 4 x 10" 4 to 1.6 1 0" 3 . The LC50 was 1.6 x 10 _ 3 M for. females. However, the LT 50 at 1.6 10 " 3 M was about 35 h. Combined results shown in Figs. 2 & 3 emphasiz the importance of food carrier in the adult feeding consumption an consequently the level of mortality due to the light-activated dy toxicity. Our data attest to the insecticidal activity of Phloxine B and Uranin mixture to Mediterranean fruit fly adults and provide fundamenta information for developing phloxine B + uranine + bait formulations fo large-scale, area-scale, area-wide spray applications. The ratio and absolute amounts of the photodye, attractant bait i commercial formulations may vary and may be readily predetermined b the practitioner skilled in the art by routine testing. It will be recognize that the photodye should be employed in an amount effective to result in significant growth inhibition or mortality rate of a test group of th target insect as compared to an untreated group. Furthermore, the amoun of attractant bait employed should be effective to attract the targe insect and stimulate it to feed on the bait. The actual effective amount of the photodye and attractant bait may vary with environment conditions such as temperature, humidity and wind, the type of vehicle carrier employed, application protocol, and stage of target inse development. Example 4 Determining Functional Relationship Betwee Concentration of Dyes and Mortality of Oriental Fruit Fly Adults Phloxine B and Uranium Mixed i/i 1% NuLure; Adult Provided Wit A Feeding Choice. Concentration-mortality relationship wa determined using the following molarity (M) concentrations of 1:1 mola mixture B and uranine in 1% NuLure: 4 x 10 " 4 , 8 x 10 "4 , 1.6 x 10" 3 . Th assay procedure was similar to the methodology described abov However, the test adults were provided with a feeding choice of tw types of food: 2 wicks with only 1% NuLure and another 2 wicks with 1 NuLure mixed with dyes. Control flies were fed with 1% NuLure. Mortalit counts were done up to 2 d. Three groups of 20 flies (by sex) were teste for each dye concentration. Adults used in the test were not starved (i.e 24 h prior to their use in the experiment, adults had sugar-protein-wat diet during the 1 2-h light phase and were provided with water only durin the 12-h dark phase). Res u lts (Figure 4) show the high toxicity (>60%) of 1 :1 molar mixture B an uranine to Oriental fruit fly adults, even in the presence of a choice food that do not contain the dyes. Example 5 Hydrolyzed Protein Bait Sprays Containing Phloxine B an Uranine for Controlling Mediterranean Fruit Fly and Orient Fruit Fly (Diptera: Tephritidae) Infestations. Insecticidal efficac of Phloxine B and Uranine mixed in hydrolyzed protein bait sprays wa assessed against an established, high density Mediterranean fruit fl population in coffee fields on Kauai, Hawaii. An experimental use perm from the State of Hawaii allowed treatment of 10 acres amidst thousand of acres of productive coffee fields. A mixture of 2.2 oz of a 1 :1 mola mixture of Phloxine B and Uranine, 1 6.23 fl oz of hydrolyzed protein ba (NuLure), and 1 5.5 gal of water was ground-sprayed for 20 application over a 3 month period on 1 0 acres of coffee, while there was no sprayin on an adjacent (control) 1 0 acres. Densities of Mediterranean fruit fly an oriental fruit fly adults were reduced by 50% in the treated field after th completion of the spraying program, with corresponding suppression o their level densities in coffee berries collected from the treated field These results were significant because the thousand of acres surroundin the 10-acre treatment plot produced hundreds of thousands of adults frui flies that were constantly immigrating into the treatment plot. Example 6 Determining Variation in Oriental Fruit Fly Adult Mortality due to Concentration of Dyes, Sex, and Time after Feeding. Phloxine B and Uranine Mixed in Aqueous Solution containing 20% Yeast Hydrolysate and 20% Fructose. Eleven concentrations, consisting of 1.25 x 10 "5 M, 2.5 x 10 *δ M, 5.0 x 10 "5 M, 1.0 x 1CT* , 2.0 x 10- 4 M, 4.0 x 10-* M, 8.0 x 10 "4 M, 1.6 x 10 "3 M, 3.2 x 10 "3 M, 6.4 x 10 '3 M, and 1.28 x 10 *2 M of 1:1 molar mixture of phloxine B (829 g per mole) and uranine (376 g per mole) wera-prepared in a stock of 20% yeast hydrolysate and 20% fructose. Six-d old adult flies confined in feeding chambers were provided with the treatment solutions saturated in cotton wicks; control flies were provided with the stock solution. Each feeding chamber had either 20 males or 20 females. Adult feeding with dye-stock mixtures commenced at 08:00 h and terminated at 12:00 h, providing a 4-h feeding duration. At the onset of feeding each feeding chamber was kept 10 cm below two hig intensity, cool fluorescent lights covered with an 80% shade cloth yielding surface light intensity of 600 lux. Following the feeding period, the shade cloth was removed and two additional fluorescent tubes were turned on, resulting in a light intensity of 18,000 lux which was then maintained throughout the remaining 12-h light phase of the experiment Upon removal of the wicks containing the treatment solutions, flies were provided with water and a diet consisting of sucrose, protein hydrolysate, and torula yeast. Mortality counts were made at the time wicks were removed and 2 h, 4 h, 6 h, 24 h, and 48 h later. The experiment followed a randomized complete block design with 7 replicates per sex-treatment combination. Results Figures 5 A and 5B show the variation in oriental fruit fly adult mortality due to concentration of dyes and time after feeding for males and females, respectively. Mortality of oriental fruit fly adults differed with dose and number of hours after termination of feeding, but was similar for males and females. Rate of kill increased with increasing concentration, with 100% mortality achieved after 2 h of light exposure at high dye concentrations (6.4 x 10 *3 M and 1.28 x 10 "2 M). Concentrations less than equal to 1.0 x 10"* failed to increase mortality beyond control levels. Example 7 Determining Relative Mortality of Oriental Fruit Flies when Exposed to D Mixed with Methyl Eugenol as compared to Methyl Eugenol alone or Methyl Eugenol + dog collar (i.e., source of the insecticide Naled). Containers with 4 - 2. cm diameter openings on the sides and holding a 2.5 cm long cotton wick holding 1.0 ml of either methyl eugenol alone (treatments 1 and 2) or a 1.28 x 10 "2 M concentrati of Phloxine B + Uranine in methyl eugenol (treatment 3) were set out in an orchard, supplemental release of irradiated oriental fruit flies. Treatment 2, additionally, had a strip of dog collar added to the interior of the container as a knock-down toxicant. On and one-half hours after the fly release, containers were collected with screen added the openings to trap all contained flies. Containers were then exposed to full dayligh and mortality counts were taken every half hour for the next two hours, after which th total number of flies was counted in each chamber. The experiment followed a randomized complete block design with 5 replicates per treatment. Results Figure 6 shows the average mortality in the three treatments over time. Ther was a high initial rate of mortality in the treatment including the dog collar strip. Two hours after exposure to full sunlight, there was greater than 98% mortality in contain to which dye had been added to the methyl eugenol wicks, compared to 100% morta in containers which had a strip of dog collar and less than 25% mortality in chambers with only methyl eugenol contained in the wicks. Example 8 Determining Whether Increase of Dye Concentration in Methyl Eugenol above 1.28 x 10 "2 M Improves the Rate of Kill of Oriental Fruit Flies. Containers with 4 - 2.5 cm diameter openings on the sides and holding a 1.8 cm long cotton wick holding 1.0 ml of either methyl eugenol alone (treatments 1 and 2) or a 1.28 x 10' 2 M (treatment 3) or 4.29 x 10 "2 M (treatment 4) concentration of Phloxine B + Uranine in methyl eugenol were set out in a citrus orchard known to have a wild population of oriental fruit flies. After two hours traps were recovered with screens added to the hole io entrap any contained flies. Wicks In treatment numbers 2 - 4 were replaced will ι water saturated wicks while wicks in treatment number 1 were not exchanged or removed. Containers were moved to full daylight and mortality counts were taken periodically over the next two hours, after which the total number of flies was counted i each container, and the number of live flies with reddened abdomens in the container which had included dye in their initial wicks. The experiment followed a randomized complete block design with 3 replicates per treatment. Results Figure 7 shows the increase in mortality overtime in the 4 treatments. By 30 minutes after the exposure of the chambers to full daylight, over 66% of the flies in th 4.29 x 10 *2 M dye treatment containers had died compared to less than 10% of those i the 1.28 x 10 "2 M dye treatment containers. By 2 hours these percentages had increased to 100% of the flies in the 4.29 x 10 "2 dye treatment containers and less than 50% of those in the 1.28 x 10 "2 M dye treatment containers. None of the flies in the 1.28 x 10 "2 dye treatment containers which continued to survive at the end of th experiment had reddened abdomens. Average mortalities in the two methyl eugeno only treatments were less than 8% and less than 20% at these two time periods, respectively. Example 9 Determining Whether Phloxine B Alone (no Added Uranine; 4.29 x 10 *2 M and 8.58 x 10 *2 M Concentrations) in Methyl Eugenol is Effective in Killing Oriental Fruit Fly Adults and Whether Rate of Kill is Greater with 8.S8 x 10" 2 M than 4.29 x 10 *2 M. Containers with 4 - 2.5 cm diameter openings on the sides and holding a 1.8 cm long cotton wick holding 1.0 ml of either methyl eugenol alone (treatments 1 and 2) or a 4.29 x 10 "2 M (treatment 3) or 8.58 x 10 "2 M (treatment 4) concentration of Phloxine B (without the addition of Uranine) in methyl eugenol were set out in a citrus orchard known to have a wild population of oriental fruit flies. After two hours traps were recovered with screens added to the holes to entrap any contained flies. Wicks in treatment numbers 2 - 4 were replaced with water saturated wicks while wicks in treatment number 1 were not exchanged or removed. Container were moved to full daylight and mortality counts were taken periodically over the next two hours, after which the total number of flies was counted in each container; during these 2-h period, the weather condition was overcast with intermittent rains. The experiment followed a randomized complete block design with 6 replicates per treatment. Results Figure 8 shows the increase in mortality over time in the 4 treatments. After 30 minutes of exposure to full daylight conditions, over 50% of the flies had died in the 8.58 x 10" 2 M dye treatment while less than 25% had died in the 4.29 x 10" 2 M dye treatment. Both control treatments averaged less than 3% mortality after 30 minutes. The difference in total mortality between the two dye treatments, however, became less over time, both reaching 100% after 2 hours of exposure, at which point both control treatments averaged less than 21% mortality.

Claims: What is Claimed is: 1. An insecticidal composition comprising: an effective amount of an attractant bait, and an effective insecticidal amount of photoactive dye. 2. The composition of claim 1 wherein said photoactive dye is selected from the group consisting of halogenated xanthene and a mixture of halogenated xanthene and uranine. 3. The composition of claim 2 which contains between about 0.001 % to about 1 0.0% by weight of said photoactive dye. 4. The composition of claim 3 further including at least one anti-foaming agent. 5. The composition of claim 4 wherein said halogenated xanthene is Phloxine B. 6. A method of eradicating or suppressing a population of Mediterranean fruit flies or oriental fruit flies and which introduces substantially minimal safety risks to humans, agriculture, and nontargeted living creatures comprising the steps of: providing the composition of claim 1 ; causing said composition to be ingested by said fruit fly populatio by spraying or providing said composition at a location where said fruit flies feed; and whereby ingestion of said photoactive dye by a Mediterranean frui fly or oriental fruit fly causes a quantity of photons to penetrate withi the body of said fly, said quantity of photons being toxic to said fruit fly, said quantity of photons further beings non-toxic to humans and said nontargeted living creatures. 7. A method for eradicating or suppressing a population o Mediterranean fruit flies or oriental fruit flies and which introduces substantially minimal safety risks to humans, agriculture, and nontargeted living creatures comprising the steps of: providing an insecticide composition which includes at least one photoactive dye; causing said composition to be ingested by said fruit fly populatio by spraying or providing said composition at a location where said frui flies feed; and whereby ingestion of said photoactive dye by a Mediterranean fruit fly or oriental fruit fly causes a quantity of photons to penetrate within the body of said fly, said quantity of photons being toxic to said fruit fly, said quantity of photons further being non-toxic to humans and said nontarget living creatures. 8. The method of claim 8 wherein said photoactive dye is selecte from the group consisting of halogenated xanthene and a mixture of halogenated xanthene and uranine. 9. The method of claim 9 wherein said halogenated xanthene is Phloxine B. 1 0. The method of claim 9 wherein said composition further includes molasses as an attractant. 1 1 . The composition of claim 1 wherein said attractant bait is molasses. 12. The composition of claim 1 wherein said attractant bait is hydrolyzed protein. 13. The composition of claim 1 wherein said attractant bait is methyl eugenol. 14. The method of claim 7 wherein said fruit flies are oriental fruit flies and wherein said composition further includes methyl eugenol as an attractant.

Inventors: CUNNINGHAM ROY T, HEINTZ JAMES R, LIQUIDO NICANOR J, SPENCER JOHN P, CUNNINGHAM, ROY, T., HEINTZ, JAMES, R., LIQUIDO, NICANOR, J., SPENCER, JOHN, P.
Applicants: PHOTODYE INT INC, US AGRICULTURE, PHOTODYE INTERNATIONAL, INC., THE UNITED STATES OF AMERICA, REPRESENTED BY THE S
Application year: 1995
Main classification: A01N25
IPC codes: A01N25, A01N43, A01N61