WO1992017062A1 - Method of identifying the avian repellant effects of a compound and method of repelling birds from materials susceptible to consumption by birds - Google Patents

Abstract

There is provided by the invention avian aversive compounds such as ortho-aminoacetophenone, 4-keto benztriazine anthranilic acid, and coniferyl benzoate. Also provided by the invention is a method for use of repellent compounds in deterring consumption of fluids, for example, non-potable fluids. Also provided by the invention is a method of identifying compounds capable of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising the steps of: a) selecting a compound, of unknown capability of repelling birds from consuming or utilizing said material, having one of the core structures (I), (II), (III), wherein R1 or R1' or R1'' is an electron donating group and R2 is an electron withdrawing group or a neutral group which group does not substantially hinder electron donation to the phenyl ring by R1. Also provided by the invention is a method for repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising applying a novel avian repellent compound identified from the method of the invention to the material in an amount effective to reduce the consumption or utilization of the material by a statistically significant amount.

Description

METHOD OF IDENTIFYING THE AVIAN REPELLENT EFFECTS OF A COMPOUND AND METHOD OF REPELLING BIRDS FROM MATERIALS SUSCEPTIBLE TO CONSUMPTION BY BIRDS

Cross Reference to Related Applications This is a continuation-in-part of application Serial

No. 679,432, filed April 2, 1991.

Field of the Invention

This invention relates to a method for repelling birds from materials such as water and edibles, and from artificial surfaces such as airport runways and parking lots, by treating the materials or artificial surfaces with avian aversive agents. The invention also relates to a method of predicting the avian aversive characteristics of certain compounds.

Background of the Invention

Regulatory constraints for operating wastewater impoundments.

Growing human populations place increasing demands on agriculture and industry. Processes from industry and agriculture often produce by-products, such as waste water, which must be stored in impoundments until it can be safely processed. While these impoundments may meet Federal and State regulations pertaining to protection of groundwater, they often pose an inherent risk to wildlife. Allen, C. , "Mitigating impacts to wildlife at FMC Gold Company's Paradise Peak Mine", McQuivey, R. , coord. Proc. Nev. wildl./mining workshop Nev. Mining Assoc. , Nev. Dept. Minerals, and Nev. Dept. Wildl. 67-71 (1990); Kay, F.R., "NDOW's role: past, present, future", ibid 18-22 (1990) . Waterfowl and other game species are often attracted to freestanding water. Should the wildlife drink from impoundments they risk death or exposure to the bioaccumulation of toxic substances, e.g. heavy metals and mutagens.

There is ample evidence to indicate that bioaccumulation of toxicants can decrease the reproductive capacity of waterfowl, and hence negatively affect wildlife populations. Ohlendorf, H.M. , et al., "Nest success, cause- specific nest failure, and hatchability of aquatic birds at selenium-contaminated Kesterson Reservoir and a reference site", Condor 91:787-796 (1989); Williams, M.L., et al., "Recruitment failure in American avocets and black-necked stilts nesting at KestersonReservoir, California, 1984-1985", Condor 91:797-802 (1989). In other cases, the actual impact of impoundments on wildlife populations may be negligible, but because of treaty concerns protection of wildlife is an important issue. For example, the Migratory Bird Treaty Act (16 U.S.C. §§ 703-711) sets zero tolerance for bird mortality. See also, Lacey Act, 18 U.S.C. § 42-44; Black Bass Act, 16 U.S.C. § 851-856; Bald Eagle Protection Act, 16 U.S.C. §§ 668- 668d; Tariff Classification Act of 1962, 19 U.S.C. § 1202, (Schedule 1 , Part 15D, Headnote 2, T.S.U.S.); Endangered Species Conservation Act of 1969, 16 U.S.C. §§ 668aa-668cc-6) . The U.S. Fish and Wildlife Service has targeted the mining and petroleum industries for enforcement in an attempt to eliminate the attractive nuisance that wastewater ponds represent to birds, and to bring these industries in compliance with the Migratory Bird Treaty Act. However, traditional hazing methods are ineffective at achieving zero mortality. (Kay, supra (1990) ; Jackson, W. B. "Bird Repelling Techniques. Pages 46-50. in R. McQuivey, coord. Proc. Nev. wildl./mining workshop. Nev. Mining Assoc. , Nev. Dept. Minerals, and Nev. Dept. Wildl (1990) . The only current commercially available effective means of preventing wildlife from using ponds is exclusion by netting. Because wastewater ponds typically range from 1 to 400 acres, this option is often impractical. For example, FMC Gold Company, spent $8 million (in netting) at the Paradise Peak Mine to exclude waterfowl; this investment resulted in reducing avian mortality from 1,548 in 1986-87 to 88 in 1988-89. Allen, C. , supra, Department of Wildlife, State of Nevada Statistics on bird mortality (1990) . The inability to reduce mortality to zero reflects the failure of netting under variable and severe weather conditions. Thus despite substantial reductions in avian mortality, the results of attempted exclosure still do not meet the requirements set forth by the U.S. Fish and Wildlife Service.

Other means of protecting wildlife are also expensive. For example, the gold/silver mining industries, in which U.S. sales were over $3.3 billion for 1989, use cyanide to extract heavy metals from ore. Because cyanide is used, the leachate impoundments are highly toxic to wildlife. Eliminating the cyanide used in the mining industries from ponds via quenching may cost about $240-400,000/year for a mid sized operation. Excluding birds from ponds until cyanide reclamation or quenching can be achieved is also costly, running between $9-13,000/acre, resulting in costs of $36- 404,000 for a range of pond sizes from 3 to 45 acres. Schroeder, M.L., "The netting of cyanide ponds at Copperstone Gold", Pages 72-81. in R. McQuivey, coord. Proc. Nev. wildl./mining workshop. Nev. Mining Assoc, Nev. Dept. Minerals, and Nev. Dept. Wildl (1990) . Further, quenching is often not desireable because cyanide can be recovered and used again. Economic figures for the petroleum industry and agriculture wastewater drainage basins are not readily available. However, wastewater negatively impacts wildlife. Successful breeding at agriculturally contaminated sites has all but ceased due to bioaccumulation of selenium in eggs. Ohlendorf, H.M. , et al., supra , (1989). The U.S. Fish and Wildlife Service is seeking methods to discourage birds from breeding at the contaminated reservoirs. Methods proposed have been as drastic as poisoning the aquatic invertebrate communities in the reservoir so as to eliminate bird food resources. To date, no method has been effective.

The above data indicate that birds are at risk when they come in contact with wastewater. It is also clear that industry and agriculture have a substantial investment to protect, and that protection can be enhanced by complying with regulatory statutes. Currently, there is a need to develop an economical alternative or ancillary strategy for keeping birds out of wastewater. Certain embodiments of the present invention relate to materials and methods for dissuading birds from drinking or otherwise utilizing industrial or agricultural wastewater. This invention discloses, inter alia, a method whereby consumption of wastewater is reduced to zero, or to levels within the toxicological tolerance of avian species.

Aviation industry and application to free-standing freshwater.

An additional area of conflict between wildlife and humans arises at airports. Blokpoel, H. , "Bird hazards to aircraft". Can. Wildl . Seirv. , Ottawa, Canada. 236pp. (1976). Many airports report numerous airstrikes with birds. In 1989, the economic losses to the U.S. military operations were on the order of $80 million. Civilian losses were reported to be a minimum of $100 million (USDA-FAA liaison office, Atlantic City) . Birds are often attracted to airports after rains because of the freestanding water which accumulates on tarmacks and runways. As is the case in mining operations, traditional hazing techniques are ineffective in that the birds are only moved from one location to another near the airport or soon become habituated to the hazing. The goal is to dissuade the birds from using the airport at all. Certain embodiments of the present invention relate to materials and methods for eliminating consumption and use of non-potable water pools formed after rain storms or after irrigation.

Background for edible additives and agricultural applications. Blackbirds and starlings (Sturnus vulgaris) can cause significant feed loss seasonally at cattle and swine operations, with the larger feedlots suffering the most damage. See e.g., Besser, J.F., et al., "Baiting starlings with DRC-1339 at a cattle feedlot", J. Wildl . Manage . 31:48-51 (1967); Palmer, T.K., "Pest bird damage control in cattle feedlots: the integrated systems approach", Proc. Vertebr. Pest. Conf . Monterey, Calif (1976); Feare, C. . , "The economics of starling damage", Econ of Dam 2:39-54 (1980). Estimates of the grain component of feed lost to birds ranges from 10-12%. Feare, C.J., et al. "Starling damage and its prevention at an open-fronted calf yard", Anim. Prod. 26:259- 265 (1978) . The risk of avian feedlot depredation to individual farmers is significant. Twenty-six percent of farmers in Tennessee reported more than negligible damage, with 6% reporting significant losses of feed to birds. Glahn, J.F., et al., "Dimethyl anthranilate as a bird repellent in livestock feed", Wildl . Soc. Bull . 17:313-320 (1989). Use of repellents to reduce consumption would be beneficial if costs of the repellent could be kept below 10% of the cost of the feed.

Birds also cause significant damage to livestock by transmitting disease. For example, over 10,000 pigs were lost to gastroenteritis during the winter of 1978-79 in one county in Nebraska. Although the total number of swine and cattle farms in not readily available for the U.S., it is clear that the potential economic loss of stock through transmissible disease is large. Certain embodiments of the present invention relate to materials and methods for reducing livestock losses due to certain avian species.

Birds also cause significant damage to crops. Currently, there is a need to develop non-lethal bird repellents to control avian crop depredation and accidental bird poisonings. Mason, et al., "Anthranilate repellency to starlings: Chemical correlates and sensory perception," J^". Wildl Manage, 53:55-64 (1989). Certain embodiments of the present invention relate to materials and methods for reducing crop losses due to certain avian species.

Sensory Biology of Birds.

Repellency in birds is substantially different than that in mammals. Szolscanyi, J. , et al., "Nociception in pigeons is not impaired by capsaicin". Pain 27:247-260 (1986) ; Mason, J.R., et al., "Exploitable characteristics of neophobia and food aversions for improvements in rodent and bird control". Pages 20-39 in D. E. Kaukienen, ed. Vertebrate pest control and management materials . Am. Soc. for Testing and Materials, Philadelphia, Pa. 315pp (1983) . Avoidance of a compound can be based on postingestional factors, e.g., toxicity, where a conditioned aversion to a sensory cue is learned. Avoidance can also be mediated via purely sensory cues. Clark, et al., "Chemical Repellency in Birds: Relationship between chemical structure and avoidance response," J Exper Zool 260: (1991). Olfaction and trigeminal chemoreception underlie the aversiveness of methyl and dimethyl anthranilate to birds (Mason, J.R. , et al., "Anthranilate repellency to starlings: chemical correlates and sensory perception", J. Wildl . Manage. 53:55-64 (1989)), suggesting that avoidance is based upon odor quality and irritation. These findings are in sharp contrast to earlier findings claiming that the limited taste capacities of birds mediated repellency. Recent findings indicate that birds are fully capable of making quantitative and qualitative odor discriminations. Mason, J.R. , et al. "Conditioned odor aversions in starlings (Sturnus vulgaris) , possibly mediated by nasotrigeminal cues". Brain Res .269:196-199 (1983); Mason, J.R. , et al., supra, (1989); Clark, L. , et al., "Olfactory discrimination of plant volatiles by the European starling", Anim. Behav. 35:227-235 (1987); Clark, L. , etal.. Sensitivity of brown-headed cowbirds to volatiles". Condor 91:922-932 (1989) ; Clark, L. , et al., "Seasonal shifts in odor acuity by starlings", J. Exp. Zool . 255:22-29 (1990); Clark, L. , "Odor detection thresholds in tree swallows and cedar waxwings". Auk 108: 177-180 (1991).

Methyl and dimethyl anthranilate (MA and DMA, respectively) are ester derivatives of anthranilic acid. MA, DMA and other ester derivatives of anthranilic acid as well as esters of phenylacetic acid, have been shown to be effective bird repellents with preferred embodiments as feed additives to deter feed loss (U.S. Patents 2,967,128 and 4,790,990) and as anti-grazing compound for geese and swans (Mason, J.R. , supra , (1989)). However, there are a number of ester derivatives of anthranilic acid, only some of which are repellent to birds. For example, linalyl anthranilate is a good repellent, whereas phenethyl anthranilate, a compound with the same molecular weight, is not repellent. Clark, et al., supra , (1991). This data suggests that isomerization is not the only important factor in identifying bird repellent compounds.

Further, DMA sprayed as a simple aqueous emulsion sprayed on food lacks sufficient taste persistency to serve as an economically attractive taste aversive agent. U.S. Patent 4,791,990. Although emulsion in a liquid other than water may increase taste persistency and thereby increase avian aversiveness, it is suggested that once evaporation of the emulsive agent occurs, taste persistency will rapidly decrease. Further, DMA is light sensitive and will degrade to a noneffective form in sunlight without adequate protection. Currently, there is a need to develop bird repellent chemicals which are capable of application in a simple emulsion. Such repellents are preferred because of the need for bird repellent chemicals that may serve as safe repellent additives to agricultural products and standing water and that might serve as safe repellents in a sprayable form.

Coniferyl benzoate, a compound found in quaking aspen (Populus tremuloides Michx) , is an important factor mediating ruffed grouse food selection. Jakubas, . & Guillon, "Coniferyl benzoate in quaking aspen a ruffed grouse feeding deterrent," J^". Chem. Ecol . 16:1077-87 (1990); Jakubas , et al., "Ruffed grouse feeding behavior and its relationship to the secondary metabolites of quaking aspen flower buds," J^". Chem. Ecol . 15:1899-1917 (1989). Currently, there is a need to identify naturally occurring compounds that would repel omnivorous birds such as starlings.

The ability to identify an ecologically sound avian repellent has numerous advantages. There is an advantage to a bird repellent that poses little or no environmental risk due to a low potential for bioaccumulation and a specific biological action. The paramount advantage is that the birds can be kept away from crops or other materials while not increasing mortality risk due to exposure to the repellent. There is a further need for bird repellent chemicals that serve as safe repellent additives to agricultural products or standing water and that might serve as safe sprayable repellents for use on crops. Such repellents are preferred because agriculture, industry and wildlife interests would be met. If used to reduce wildlife hazards associated with formulated agricultural chemicals, the ecologically sound avian repellant also could have an enormous economic impact, particularly on major American chemical producers, and the farmers who rely on the continued availability of the producer's agricultural products.

Summary of the Invention >

There is provided by this invention several novel avian repellents. There is further provided by this invention a novel method of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising applying a compound to said material, wherein said compound is alpha-aminoacetophenone, ortho- aminoacetophenone, meta-aminoacetophenone, para- aminoacetophenone, ortho-hydroxyacetophenone , ortho- methoxyacetophenone , eta-methoxyacetophenone, para- methoxyace ophenone, anthranilic acid, 3-aminobenzoic acid, 4-aminobenzoic acid, 4-ketobenztriazine, coniferyl benzoate. cinnamyl benzoate, 3-4 dimethoxycinnamyl benzoate, coniferyl alcohol, cinnamyl alcohol, or 3-4 dimethoxycinnamyl alcohol, in an amount effective to reduce the consumption or utilization of said material by said birds by a statistically significant amount.

There is further provided by this invention a novel method of repelling birds from consuming or utilizing aquatic habitats, including but not limited to non-potable water, comprising applying an ester of anthranilic acid or mixtures thereof to said aquatic habitat in an amount effective to reduce the amount of consumption or utilization of said aquatic habitat by said birds by a statistically significant amount.

There is further provided by this invention a novel method of identifying compounds capable of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising selecting a compound, of unknown capability of repelling birds from consuming or utilizing said material, having one of the following core structures;

wherein; R₁ or R,' or R.," is an electron donating group (R, being ortho, meta or para) and R₂ is an electron withdrawing group or a neutral group which group does not substantially hinder electron donation to the phenyl ring by R^* and optionally testing said compound to confirm the capability of said compound to repel birds from consuming or utilizing said material.

There is further provided by this invention a novel method for repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising applying a novel avian repellent compound identified according to the method described herein to said material in an amount effective to reduce the amount of consumption or utilization of said material by a statistically significant amount.

Despite increasing demand, few non-lethal chemicals

(i.e., repellents) are available for the control of avian depredation and nuisance problems. Until now the small number of compounds known in the art to be capable of repelling birds from consuming or utilizing materials otherwise susceptible to consumption by birds, were identified serendipitously without recognition of the underlying principles taught herein. The invention has considerable value in that it represents an empirical addition to a restricted body of chemicals that effectively repel birds from consuming or utilizing materials susceptible to consumption or utilization by birds. Therefore, the present invention, by providing additional novel bird repellent compounds, represents a significant contribution to improved agricultural technology, and fulfills a technological need.

Currently, there are no chemicals commercially available to prevent the accidental ingestion of pelleted agricultural chemicals, treated seeds, agricultural wastewater, or the toxic solutions found in industrial evaporating ponds. Shah, et al., "Prediction of Avian Repellency from Chemical Structure: The aversiveness of vanillin, vanillyl alcohol, and veratryl alcohol," Pesticide Biochem and Physiol 40:169-175 (1991). The present invention provides a method of chemically repelling birds frommaterials including but not limited to the above identified materials. Additionally, if used to reduce the non-target hazards associatedwithmammalian toxicants and agrichemicals, the invention could have a substantial positive environmental impact. If used to reduce wildlife hazards associated with formulated agricultural chemicals, the invention could have an enormous economic impact, particularly on major American chemical producers, and the farmers who rely on the continued availability of the producer's agricultural products. Brief Description of the Drawings

Figure 1. Laboratory drinking trials for European starlings. N=9 birds per experimental group. During testing birds were given: deionized, distilled water (H₂0) , water from cyanide holding ponds with cyanide removed (Pond) , deionized, distilled water treated with MA (MA-H20) , and pond water treated with MA (MA-Pond) .

Figure 2. Ohio pen trials. Frequency of drinking behavior for treated (MA) and untreated (C) pools for ducks and gulls. For ducks, the data reflect the combined frequency for both formulations of MA.

Figure 3. Ohio pen trials. Frequency of entry into treated (MA) and untreated (C) pools for ducks and gulls. For ducks, the data depicted reflect the combined frequency for both formulations of MA.

Figures 4a - 4i illustrate the structural similarities of acetophenones. Arrows indicate donation of lone electron pairs. Figure 4a is ortho-aminoacetophenone, Figure 4b is para-aminoacetophenone and Figure 4c is eta- aminoacetophenone. Figure 4d is ortho-methoxyacetophenone, Figure 4e is para-methoxyacetophenone and Figure 4f is meta- methoxyacetophenone. Figure 4g is orthos-hydroxyacetophenone, Figure 4h is para-hydroxyacetophenone and Figure 4i is meta- hydroxyacetophenone. Figures 5a - 5i illustrate the mean dose response curves for acetophenones recorded during the 6 hour drinking trials of European starlings. Vertical lines depict + one standard error. Sample size N=4/concentration group. Figure 5a is 2-HAP. Figure 5b is 3-HAP. Figure 5c is 4-HAP. Figure 5d is 2-M0AP. Figure 5e is 3-M0AP. Figure 5f is 4-M0AP. Figure 5g is 0AAP. Figure 5h is MAAP. Figure 5i is PAAP.

Figures 6a - 6b illustrate the mean dose response curves and structures for benzoic (Figure 6a) and 2- hydroxybenzoic acid (Figure 6b) . Vertical lines depict ± one standard error. Sample size N=6/concentration group.

Figures 7a - 7c illustrate the mean dose response curves and structures for 2-methoxybenzoic acid (Figure 7a) , 3-methoxybenzoic acid (Figure 7b) and 4-methoxybenzoic acid (Figure 7c) . Vertical lines depict + one standard error. Sample size N=6/concentration group.

Figures 8a - 8c illustrate the mean dose response curves and structures for anthranilic acid (Figure 8a) , 3- aminobenzoic acid (Figure 8b) and 4-aminobenzoic acid (Figure

8c) . Vertical lines depict ± one standard error. Sample size

N=6/concentration group.

Figures 9a - 9c illustrate the mean dose response curves and structures for phenyl rings with attached amides and 5-NAA. Figure 9a is 5-nitroanthranilic acid. Figure 9b is anthranilamide. Figure 9c is benzamide. Vertical lines depict + one standard error. Sample size N=6/concentration group. Figures 10a - 10c illustrate the dose response curves for phenyl rings with attached heterocyclic rings and o-carboethoxybenzene sulfonamide. Figure 10a is orthocarboethoxybenze sulfonamide. Figure 10b is isatoic anhydride. Figure 10c is 4-ketobenztriazine. Figures 11a - lib. Figure 11a illustrates the mean consumption (left) and preference ratios (right) from 1-cup tests with ortho-aminoacetophenone. Figure lib illustrates the mean consumption (left) and preference ratios (right) from 2-cup tests with ortho-aminoacetophenone. Preference ratios were generated by dividing consumption from a random cup (pre- treatment) or the treated cup (treatment) by total consumption. A ratio of 1.0 indicates total preference; 0.0, total rejection; and 0.5, indifference for feed in the randomly selected or the treated cup. Capped vertical lines represent standard errors of the means.

Figures 12a - 12b. Figure 12a illustrates the mean consumption (left) and preference ratios (right) from 1-cup tests with para-aminoacetophenone. Figure 12b illustrates the mean consumption (left) and preference ratios (right) from 2- cup tests with para-aminoacetophenone. Capped vertical lines represent standard errors of the means. Figures 13a - 13b. Figure 13a illustrates the mean consumption (left) and preference ratios (right) from 1-cup tests with meta-aminoacetophenone. Figure 13b illustrates the mean consumption (left) and preference ratios (right) from 2- cup tests with meta-aminoacetophenone. Capped vertical lines represent standard errors of the means.

Figures 14a - 14b. Figure 14a illustrates the mean consumption (left) and preference ratios (right) from 1-cup tests with alpha-aminoacetophenone. Figure 14b illustrates the mean consumption (left) and preference ratios (right) from 2- cup tests with alpha-aminoacetophenone. Capped vertical lines represent standard errors of the means.

Figure 15 illustrates the mean consumption of acetophenone treatment minus pretreatment food consumption levels (one cup difference scores) . Negative values indicate repellency, positive values depict preference and 0, indifference.

Figures 16a - 16c. Figure 16a illustrates the ranking of relative water consumption as a function of isomeric position of the electron donating group. Ortho and para isomers are capable of resonance of lone electron pairs, the meta isomers are not. Consumption is relative to consumption of water controls. Figure 16b illustrates the ranking of relative water consumption as a function of basicity of the molecule. Amino substituents are the most basic; hydroxy substituents are the least basic. Figure 16c illustrates the ranking of relative water consumption as a function of the presence (YES) or absence (NO) of intramolecular hydrogen (H) bonding. Figures 17a - 17g. Chemical structures of coniferyl benzoate (Figure 17a) , coniferyl alcohol (Figure 17b) , cinnamyl benzoate (Figure 17c) , cinnamyl alcohol (Figure 17d) , 3,4-dimethoxycinnamyl benzoate (Figure 17e) , 3,4- dimethoxycinnamyl alcohol (Figure 17f) , and benzoic acid (Figure 17g) .

Figures 18a - I8g illustrate the mean treatment consumption of treated and control feed by starlings in 2-cup tests. Capped vertical bars represent standard errors of the mean. Figure 18a is benzoic acid. Figure 18b is cinnamyl benzoate. Figure 18c is cinnamyl alcohol. Figure 18d is 3,4- dimethoxycinnamyl alcohol. Figure 18e is coniferyl benzoate. Figure 18f is coniferyl alcohol. Figure 18g is 3,4- dimethoxycinnamyl benzoate.

Figure 19. Mean 2-cup preference ratios, calculated by dividing treated feed consumption by total (treated and control) consumption. A preference ratio of 1.0 indicates absolute preference for treated feed, a ratio of 0.0 indicates absolute rejection of treated feed, and a ratio of 0.5 indicates indifference between treated and control feed.

Figures 20a - 2Oh illustrate the mean pretreat ent and treatment consumption by starlings in 1-cup tests. Capped vertical bars represent standard errors of the means. Figure 20a is benzoic acid. Figure 20b is cinnamyl benzoate. Figure 20c is cinnamyl alcohol. Figure 20d is 3,4-dimethoxycinnamyl alcohol. Figure 20e is coniferyl benzoate. Figure 20f is coniferyl alcohol. Figure 20g is 3,4-dimethoxycinnamyl benzoate. Figure 20h is benzoin Siam.

Detailed Description of the Invention

Methods are herein provided of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds. As used herein, the term "birds" refers to members of the class "Aves".

The compounds disclosed herein are available from a variety of commercial sources such as Aldrich Chemical Co. ,

Milwaukee, Wisconsin. Although not wishing to be bound by any particular mechanism of action, it is believed that birds are repelled in the method of the invention through olfaction and nasal trigeminal chemoreception, i.e. repellency may be based on odor quality and irritation. Those skilled in the art will recognize bird utilizations of materials which would benefit from the method of the invention. Essentially utilization refers to contact by the bird with said material. Such utilizations that may be repelled include e.g. , bill dipping in contaminated waters, landings on a variety of hazardous surfaces such as runways and parking lots.

In the practice of the present invention, the compounds disclosed herein may be applied to the material to which the bird is to be repelled in any suitable manner. For example, liquid carriers may be employed and the repellent may be sprayed on the material. See e. g. U.S. Patent 2,967,128 which patent is incorporated by reference as if fully set forth herein. In another embodiment, the repellent can be at least partially trapped in a solid vehicle to improve its persistency such as disclosed in U.S. Patent 4,790,990 which patent is incorporated by reference as if fully set forth herein. The vehicle can be a modified starch, oil or polymer which microencapsulates the aversive agent.

"Materials", to which birds are to be repelled from consuming or utilizing, as used herein refers to materials otherwise susceptible to consumption or utilization by birds. Examples include edible and non-edible materials such as water, feeds, waste waters, seeds and agrichemicals such as pesticides and herbicides.

A material of particular interest for the method of the invention includes non-potable liquids such as industrial or agricultural waste water, mine tailing ponds, and freestanding water on artificial surfaces like airport runways and parking lots. "Non-potable refers to liquids or aquatic habitats wherein said liquid is not fit to drink but may be consumed to the detriment of the bird. The repellant can be dispersed in the liquid to reduce the amount of liquid consumed by avian species, and to reduce the likelihood that birds will rest or swim in the liquid. The repellant may be incorporated into freestanding water, industrial or agricultural wastewater or fluid toxic containment ponds to reduce the likelihood that birds will drink, rest or swim in the liquid, resulting in reduced mortality and morbidity to birds. Without recourse to water resources, birds will vacate the treated area.

This invention also discloses a repellant to materials such as feed or pesticides, such as ortho- aminoacetophenone, 4-ketobenztriazine or anthranilic acid. In one embodiment, the repellent is at least partially trapped in a solid vehicle to improve its persistency. The vehicle is a modified starch, oil or polymer which encapsulates, emulsifies or substantially uniformly disperses the aversive agent. The disclosed additive is dispersed into solids consumed by avian species to reduce the likelihood that they will eat the treated edible. The general field of application is bird damage control, with the additive to be used in application to fruit and grain crops, livestock feeds, non- food crops (e.g., clover, turf) or as a protective seed coating. The additive can also be used to reduce non-target hazards associated with mammalian toxicant and agrichemicals. The invention could have a substantial positive environmental impact. In the preferred embodiments, the effective amount of the compound is that amount sufficient to reduce the consumption or utilization of said material by said birds to at least about 50 percent (and most preferably at least about 90 percent) over the amount of said material which would otherwise be consumed or utilized by said birds without said compound applied.

In another embodiment of the invention a method of identifying compounds capable of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds is provided. A compound, of unknown capability of repelling birds from consuming or utilizing said material, having one of the core structures defined herein (See formula (1) ) is selected wherein; , or R.,' or R.," is an electron donating group; and the compound is optionally tested to confirm the capability of said compound to repel birds from consuming or utilizing said material. Electron donating groups are well known to those skilled in the art and a variety of suitable examples are shown below and include amine, o-lower alkyl, N-lower alkyl, and N-di lower alkyl. Specific examples would include NH₂, NHCH₃, NC₂H₆, NH0C₂H₅ and 0CH₃.

Methods to test compounds to confirm their ability to repel are known to those in the art and several are detailed in the Example section below. Briefly, one suitable test comprises comparing the amount of material consumed or utilized by a bird with the amount of material consumed or utilized by a bird comprising an avian repellant amount of said compound dispersed therein in a one choice test under controlled conditions whereby a statistically significant decrease in consumption of the material with said compound confirms said compounds ability to repel birds from consuming or utilizing materials susceptible to consumption or utilization by birds. Statistical significance may be judged by appropriate statistical tests, including but not limited to a t-statistic or analysis of variance, relative to a plain water control at a probability level of 0.05 or less.

An "avian repellent amount" suitable for use in the test can be easily determined by one skilled in the art, e.g. , 1 to 0.001% v/v or w/v. For example, one can compare the amounts of other known repellents used on the same type of material to be tested. Generally, testing to confirm repellency involves a series of concentrations to be tested to determine optimal amounts for a variety of factors.

With respect to the substituents on the phenyl ring, it is preferred that R₂ is an electron withdrawing group (EWG) and/or R₂ is a group capable of forming a hydrogen bonded ring structure with an electron donating group, R.,. Both types of R₂ groups are well known to those in the art and a variety of examples of such groups are shown herein and include lower acyl, carboxylic acids, esters and nitro groups as examples of electron withdrawing groups. The other type includes groups like CH₂ and CH₂NH₂. Specific examples for R₂ include COCH₃, COOCH₃, COOC₂H₅, COOC₂H₄-phenyl, COO-linalyl, N0₂, CH₂0H and C0CH₂NH₂; Thus, R₂ can be of two forms, an EWG or not. If it is an EWG, then in association with an R, at positions 2 and 4 resonance can occur and this contributes to repellency. R₂ may also contribute to repellency even if it is not an EWG, however it should form a hydrogen bonded ring structure with R_t, e.g., 2-aminobenzyl alcohol; or an electron donating group within itself, e.g, alpha-aminoacetophenone. And it is preferred that the substituents on R₂ are of the type that do not substantially prevent R_t from donating electrons to the phenyl ring, e.g., phenethyl anthranilate (probable steric effects) or isotoic anhydride (electronic effects) .

Additionally, it is preferred that the electron donating groups, R, or R₁' or R.,", contribute electrons to the phenyl ring, most preferably in position 2, 3 or 4 with respect to R₂.

It is preferred that the electron donating group is basic. Additionally it is preferred that there is intramolecular hydrogen bonding between R, and R₂ when R₁ is in position 2 relative to R₂, e.g., ortho-aminoacetophenone or

2-hydroxyacetophenone.

R₁ and R₂ may comprise a heterocyclic ring attached to the phenyl ring such as 4-keto benztriazine.

Identification of additional novel avian repellents through the method of this invention has implications for the development of commercially viable, ecologically sound, nonlethal bird repellents. Isomers (ortho, meta, para) and moieties (amino, hydroxy, methoxy) of acetophenones were tested for their effectiveness as avian aversive agents to better understand the nature of repellency in birds.

Chemically, basicity of a substituted phenyl ring, as defined by the electron donating substituent, is believed to be an important feature influencing repellency; i.e., more basic substituents result in more potent repellents. Isomeric position of the electron donating substituent which leads to resonance of lone pairs of electrons is also an important feature of repellency; i.e., repellency is enhanced when electron donating substituents are in the ortho and para positions. An ancillary contributory factor enhancing repellency is believed to be the presence of intramolecular hydrogen bonds. Combining these features appears to improve repellency.

As stated, the strength of repellency is believed to decrease as a function of the positional isomer, e.g. ortho isomers are better repellents than para isomers, which in turn are better than meta isomers, suggesting that electron donation by resonance is an important feature of repellency. The ortho and para isomers are in direct resonance with the carbonyl group, whereas the meta isomer is not (Figure 16a) . The meta isomers will actually withdraw the electrons from the phenyl ring through the inductive effect. As stated, it was found that basicity (and/or electron donating ability of the substituent group) is an important factor controlling repellency (Figure 16b) . The relative basicity for the compounds is greatest for amino substitutions and least for hydroxy substitutions, a ranking concordant with the data on strength of repellency.

Interaction effects are believed to be important. For example, a very effective repellent is ortho- aminoacetophenone (0AAP) . This molecule is very basic and has its amino group in the ortho position. As a consequence, the amino group is in a position to form an intramolecular hydrogen bond with the electron withdrawing group, suggesting an ancillary contributory role to repellency of such bonding. Intramolecular hydrogen bonding is not believed to be crucial to repellency because the basic para and meta isomers (4AAP, 4M0AP; and 3AAP, 3M0AP, respectively) are effective repellents. Basicity, and not resonance, may be the most important feature of repellency. The relatively acidic para isomer of hydroxy acetophenone, which is capable of resonance, is an effective repellent. Only when the substituent is in the ortho position does this hydroxy acetophenone become repellent. This suggests that intramolecular hydrogen bonding interacts with resonance to overcome the negative effects of the relative acidity on repellency. Abbreviations: Compound

MA: methyl anthranilate OAAP: ortho-aminoacetophenone

MAAP: meta-aminoacetophenone

PAAP: para-aminoacetophenone

AAAP: alpha-aminoacetophenone

2-HAP: ortho-hydroxyacetophenone 3-HAP: meta-hydroxyacetophenone

4-HAP: para-hydroxyacetophenone

2-MOAP: ortho-methoxyacetophenone

3-MOAP: meta-methoxyacetophenone

4-M0AP: para-methoxyacetophenone BA: benzoic acid

5-NAA: 5 nitroanthranilic acid

ANAM: anthrani1amide

BAM: benzamide

OCEBS: ortho-cabroethoxybenzene sulfonamide IA: isatoic anhydride

4-KBT: 4-Keto benztriazine

SA: salicylic acid (2 hydroxybenzoic acid)

4ABA: 4-aminobenzoic acid

3ABA: 3-aminobenzoic acid ANA: anthranilic acid (2 amino benzoic acid)

PAA: para-anisic acid (4 methoxy benzoic acid)

MAA: meta-anisic acid (3 methoxy benzoic acid)

OAA: ortho-anisic acid (2 methoxy benzoic acid) EXAMPLES DRINKING TESTS Example 1. Methyl Anthranilate as a water repellent.

Previous work has shown MA and other ester derivatives of anthranilic acid to be effective bird repellents in the embodiment of feed or pesticide additives and topical applications to turf. See e. g. U.S. Patent No. 2,967,128. The application of MA to wastewater is not apparent from previous work in this area, even though laboratory data show that birds decrease consumption of deionized, distilled water treated with MA. The present experiment surprisingly demonstrates that waste water derived from pond tailings is actually preferred relative to deionized distilled water by birds, and that treatment with MA synergistically decreases co:.sumption beyond what one would expect based on treatment of deionized, distilled water.

Example la. Wastewater.

Procedure .

Birds.—Adult European starlings (Sturnus vulgariε) were decoy-trapped in rural New Jersey and transported to the laboratory. Upon arrival, the birds were individually caged (61 x 36 x 41 cm) under a 12:12 light:dark: cycle with light onset at 0700 h. Food was available ad libitum. Before experiments began, the birds were permitted free access to tap water.

Chemicals.—MA (CAS #134-20-3) was obtained from PMC, Specialties, Cincinnati, Ohio. Wastewater from cyanide extraction of gold was obtained from Goldfield Mines, Golconda, Nevada. One-choice Tests .—Starlings were given 3 days of pretreatment during which water consumption was measured for 6 hr. At the end of this period, individuals whose variance about the 3 day mean consumption was greater than + one standard deviation of the population variance were excluded from the trials. Those birds with stable daily water consumption were ranked according to mean water consumption and assigned to treatment groups. The bird with the highest water consumption was assigned to the deionized, distilled water treatment group, the bird with the second highest consumption was assigned to the leachate pond-water treatment, the bird with the third highest consumption was assigned to the MA+deionized, distilled water treatment and the bird with the fourth highest consumption was assigned to the MA+leachate pond-water group, and so forth, until all birds were assigned to a group. (MA concentration 0.5% v/v) After assignment to a treatment group a 1 day drinking trial began. Birds had free access to feed and tap water during the night. Beginning at 0930, the water was replaced with preassigned concentrations of chemicals and consumption was recorded every 2 hours for the next 6 hours. After the test, birds were given free access to tap water. Consumption of tap water was monitored overnight and that of deionized, distilled water the next day, and these values were compared with pretreatment drinking to determine whether consumption had returned to normal. There were no mortalities, and at the end of all experiments birds were released to the wild.

Analysis.—There were no differences of mean consumption of deionized, distilled water among groups during the pretreatment period. All data were found to be homogeneous unless otherwise noted. Data were analyzed using a one-way analysis of variance (ANOVA) with the main effect as treatment group. Post-hoc differences were determined using a Scheffe's test.

Results.—There was a significant treatment effect (Figure 1, F=23.8, df=3,68, P < 0.001). All treatments differed from one another at the P < 0.05 level. Birds consumed significantly more pond-water than any other type of water. Next, birds consumed high levels of deionized, distilled Water. Addition of MA to the deionized, distilled water reduced consumption of water to one-half the untreated level. Surprisingly, addition of MA to wastewater decreased consumption to levels statistically indistinguishable from zero consumption (£ < 0.05).

Example lb. Fresh Water Field Trials.

Gulls, waterfowl and other species frequently flock to temporary pools of fresh water at airports after heavy rains, creating safety hazards for aircraft (Blokpoel, H. , supra , (1976)). Development of environmentally safe chemical formulations that can be added to water to repel birds has wide utility. Procedure.

Birds.—Sixteen mallards (Anas platyrhincus) and 16 ring-billed gulls (Larus delawarensis) were funnel or rocket trapped at Sandusky, Ohio and kept two birds/species to a pen. Pens were 8 x 4 m corrals, each with an attached 2.5 x 2.5 x 2.5m shaded holding pen. Pens were set up on mowed grass in an area isolated from human disturbance at the Ohio Research Station of the U.S.D.A./A.P.H.I.S./S. & T. (Plum Brook Station), Erie County, Ohio. Each corral had 2, 0.8m diameter or 2, 1.0 m diameter plastic pools filled with 40L or 90L, of water (10-12cm deep) , respectively. For each of 2 days, 2 mallards, with primary feathers pulled on 1 wing to prevent flying, were placed in each holding pen and released daily for 9 hrs into the corral to acclimate to the test condition. Each corral contained a pan with cracked corn, millet and commercial duck food.

Treatments .—On test day 1 at 0800, 2 formulations of MA embedded in a polymer were applied to fresh water (0.5% w/v) in a randomly selected pool in each corral. Formulation 1 contained 16% MA and was applied to the pool as 1 part formulation to 200 parts water, yielding 0.08% concentration of active ingredient in the pool. Formulation 2 consisted of 64% MA applied to the pool as 1 part formulation to 200 parts water, yielding a concentration of 0.32% active ingredient. Water depth was measured to the nearest ml and the 2 mallards were released into the corral. One of 4 observers (2 corrals per observer) watched each corral for 12020-sec intervals (40 min total) over the next 2 hours. The observer recorded the number of mallards in each pool (pool use) during each 20-sec interval and the total number of times a bill touched the water (i.e. drinking or bathing activity) for each pool. At 1600 the water depth was remeasured and the mallards were returned to their holding pen where they were provided with food, but no water. This routine was maintained on days 2, 3, and 4. The mallards were kept in their holding pens on day 5 (with drinking water and food) . On day 6, they were released into corrals with only the MA-treated pool available. The birds were observed as before and the experiment was then terminated.

For the gull experiment the methods were the same as for mallards, except that the tests took place in the holding pens (the gulls could fly) and only 1 formulation (#2) of MA was used. The gulls were fed fresh fish daily. Four pens were used.

Analysis.—Data were analyzed using a repeated measures analysis of variance with days as the repeated measure.

.Results.—Both formulations of MA were effective in the duck experiment at keeping mallards from swimming, or bathing in MA-treated pools (Figure 3, F=46.1, P<0.001 and

F=12.5, P<0.01, df=l,3 for formulations 1 and 2, respectively) . Both formulations were also effective at inhibiting drinking and bill dipping behavior by ducks (Figure

2, F=47.1, P<0.01 and F=42.9, P<0.01, df 1,3 for formulations

1 and 2, respectively). During the 4 days of the 2-choice test 98.5% of the entries into pools by ducks and 96.1% of bill contacts were in the untreated pools. In the 1-choice test, where ducks were only exposed to pools treated with MA, use of the pools for entries and drinking was restricted to

96.2% and 91.2% relative to pretreatment levels respectively.

Repellency was even more pronounced in the gull experiments (Figures 2 and 3) . During the 2-choice tests for formulation 2, over 99% of entries and bill contacts were in the untreated pools (F=61.2, P<0.01 and F=55.7, P<0.01, df=l,3 for swimming and bill dips, respectively) . For the 1-choice tests, only a single incidence of pool use and 83 bill contacts were recorded compared with a daily means of 38.8 and 552.9 for fresh water during the previous four days. Discussions.—The dramatic reduction of water consumption for the MA-wastewater treatment group in Example la indicates a synergism between MA and the pond water. Because MA is an ester it is unstable under a variety of conditions, it readily cleaves into anthranilic acid. Anthranilic acid is also a bird repellent. It is believed that the intrinsic repellent properties of anthranilic acid and MA and the sensory aspect of MA, combine to act as a potent repellent in an aqueous medium. The enhanced repellency is not believed likely to be due to interactions with the taste of wastewater.

The concentrations of MA in the formulations in Example lb corresponded to 0.08 and 0.32%. These values are substantially lower than effective dosages reported for feeding trials in the laboratory. This may be due, in part, to the better access to receptors when repellent is in an aqueous medium. It may also be due to a combination of the slow release of MA from the matrix which would contribute to persistency of the volatile cue and the breakdown of MA into water soluble anthranilic acid. These tests demonstrate the applicability of laboratory data to water repellency in field situations. Example 2. Acetophenones

Examination of isomeric configurations of aminoacetophenones permitted inferences about the structural features of the molecule and repellency (Figures 4a - 4i) . When the electron donating amino group is in the ortho position, intramolecular hydrogen bonding between the carbonyl and amino group is possible. Electron sharing with the benzene ring is possible through resonance. When the amino group is in the meta position, no intramolecular hydrogen bonding is possible and electron sharing is primarily through induction. When the amino group is in the para position, there is still no intramolecular hydrogen bonding, but electron sharing is possible through resonance.

Methoxyacetophenones were selected because the methoxy group is less basic than the amino group and no intra- molecular hydrogen bonding is possible for this molecule. Resonance effects are similar to the aminoacetophenones for each of the isomers. Hydroxyacetophenones were selected because they are the least basic of the acetophenone moiety series, but allow intramolecular hydrogen bonding when in the ortho position. The comparison of these moieties demonstrated the relative importance of intra-molecular hydrogen bonding, basicity and resonance for avian repellency.

All isomers of aminoacetophenone (ortho- CAS# 551- 93-9, meta- CAS # 99-03-6, para- CAS #99-92-3), hydroxyacetophenone (ortho- CAS #118-93-4, meta- CAS #121-71- 1, para- CAS #99-93-4) and methoxyacetophenone (ortho- CAS #4079-52-1, meta- CAS #586-37-8, para- CAS #100-06-1) were obtained from Aldrich Chemical Company, Milwaukee, Wisconsin. Because acetophenones are generally insoluble in water, each compound was mixed in water under low heat to yield saturated emulsions with concentrations at 0.5% (weight or volume/volume) . Lower concentrations were established by serial dilutions to yield: 0.25, 0.125, 0.0625, and 0.0313 %. Procedure . One-choice Tests.—Starlings were given 3 days of pretreatment during which water consumption was measured for 6 hr. At the end of this period, individuals whose variance about the 3 day mean consumption was greater than + one standard deviation of the population variance were excluded from the trials. Those birds with stable daily water consumption were ranked according to mean water consumption and assigned to treatment groups. The bird with the highest water consumption was assigned to the 0.5% treatment group, the bird with the second highest consumption was assigned to the 0.25 % treatment group, and so forth, until all birds were assigned to a group. After assignment to a treatment group a 1 day drinking trial began. Birds had free access to feed and tap water during the night. Beginning at 0930, the water was replaced with preassigned concentrations of chemicals and consumption was recorded every 2 hours for the next 6 hours. After the test, birds were given free access to tap water. Consumption of tap water was monitored overnight and the next day, and these values were compared with pretreatment drinking to determine whether consumption had returned to normal. If this condition was met, the birds were tested with the next compound, with groups of birds receiving a different concentration of compound as determined by a counter-balanced predetermined schedule. Naive birds were not used for each test because it was not practical to capture the 324 starlings required for all experiments. If an individual's inter- experiment water consumption was within + 1 standard error of its pretreatment value the bird was used in the next experiment. In addition, birds were checked for health condition after each experiment, e.g. piloerection. There were no mortalities, and at the end of all experiments birds were released to the wild.

Analyses.—To test for carry over-effects due to treatment, the inter- treatment tap water consumption of all birds using a repeated measures, 1-way analysis of variance (anova) was examined. Each post-treatment day's water consumption by partition, using an a priori contrast with pretreatment water consumption was compared. Each isomeric moiety was considered separately.

Two a priori hypotheses about consumption of treated water were tested. (1) Did consumption of treated water differ from a theoretical value of zero consumption? This information is useful because there may be times when a bird must be repelled from potentially lethal toxic waste water, e.g. cyanide ponds resulting from precious metal extraction in the gold mining industry. McQuivey, R. , Coord. Proc. Nev. wildl./mining workshop. Nev. Mining As (soc, Nev. Dept. Minerals, and Nev. Dept. Wildl. (1990). The analysis required a slight modification in calculation of the treatment sums of squares, where the grand mean was replaced by zero and the degrees of freedom (df) reflected the number of treatments considered in the experiment (i.e. k=6) . Estimates of the error term remained the same as in a standard anova. Post-hoc comparisons were made using a modification of Dunnett's t-test (1959) , again using a theoretical value of zero rather than the mean, and comparing the resulting t to critical values in Dunnett's calculated distribution, with P set at < 0.05. (2) Did mean water consumption differ among the treatment (i.e., concentration) groups? An 1-way anova was used to compare group means and a Scheffe's post-hoc test was used to identify significant (P<0.05) differences among means.

Results.—There were no post-treatment changes in consumption of tap water caused by any of the treatments (P>0.05). Further, no bird showed signs of illness following consumption of treated water. No bird was excluded from experiments once testing began, i.e., consumption of deionized, distilled water during inter experiment intervals remained stable relative to pretreatment water consumption. As evidenced from the dose-response curves, some compounds were more effective at inhibiting water consumption (Figures 5a - 5i) . Two compounds, 3-HAP and 4-HAP were totally ineffective as repellents (Table 1) . The remaining compounds showed differences amongconcentrations tested, with 0AAP showing the strongest signs of activity (Table 1) . Overall, the aminoacetophenones inhibited water consumption the most, with methoxyacetophenones and the ortho position of hydroxyacetophenone having less of an effect relative to controls. The ortho positions within each moiety showed the strongest sign of repellency followed by the para and meta positions.

There may be circumstances when birds must be kept from drinking any water, e.g. toxic waste water impoundments. Therefore it is of interest to compare consumption against a theoretical value of zero consumption. Not all compounds were equally effective at repelling birds from drinking water (Figures 5a - 5i) . Two compounds, 3-HAP and 4-HAP were ineffective as repellents. OAAP showed absolute repellency over the broadest range of concentrations tested.

Table 1 Differences in water consumption among concentrations tested

P^b Post-hoc Scheffe test^c .06 .50 .13 .03 .25

2HAP 5.32

3M0AP 3.74

4M0AP 10.89

^■ F value from a one-way anova.

£ probability associated with the anova that consumption across all concentration groups was equal. Rejection of the hypotheses occurred when P<0.05. ^c Post-hoc test of significant differences (P<0.05) among concentration groups (values listed in Table) . Similarities for group consumption for concentration tested are connected by lines.

Example 3. Benzoic Acids

Benzoic acids were chosen to examine the effects of increasing the acidity of molecules by replacing the carbonyl electron withdrawing group attached to the benzene ring with a carboxylic acid. The amino substitution for the electron donating group was chosen for examination because it represented a highly basic substituent. As was the case for acetophenone moieties, the ortho position in anthranilic acid is capable of intramolecular hydrogen bonding, while the meta and para isomers are not. Electron sharing with the benzene ring for these isomers is through resonance for the ortho and para isomers and induction for the meta isomer. The discovery that anthranilic acid (2-aminobenzoic acid) was a good repellent led to speculation that the acidity of the withdrawing group was not a primary feature governing repellency. The methoxy and hydroxy moieties were chosen because they represented a less basic electron donating group than the amino substitution. 5-nitro anthranilic acid was selected to increase the electron withdrawal from the phenyl ring. In this case intramolecular hydrogen bonding is still possible between the amino and carboxyl groups, though the electron sharing via resonance is more uniformly spread over the entire molecule.

By adding a nitro group to anthranilic acid (an otherwise good repellent) , the addition of a strong withdrawing group was tested as to how it affected repellency

(e.g. 5-nitro anthranilic acid) . Examination of anthranilamide allowed testing of the effect of internally compensating the electron withdrawal capacity of the carboxyl group on repellency. The internal compensation for withdrawing capacity allows the donating amino group to contribute electrons to the phenyl ring. Examination of benzamide showed how internal compensation of the withdrawal groups and elimination of the donation group effects repellency. In this case, resonance within the phenyl ring is unaffected and the ring is relatively electron poor when compared to anthranilamide.

Previous work showed that esters of benzoic acids are good repellents. The findings herein indicated that planarity of the pi cloud formation is also important for repellency. Examination of o-carboethoxybenzene sulfonamide showed how distortion of the pi cloud affects repellency. Isatoic anhydride was selected because the rigid planar structure is maintained via covalent bonds but the hetero ring is strongly withdrawing, thus making the phenyl ring electron poor. 4- ketobenztriazine was selected because it maintains a planar structure in the hetero ring, but donates electrons to the phenyl ring.

All isomers of anthranilic acid (ortho- CAS #118-92-3, meta-CAS #99-05-8, para- CAS #150-13-0), anisic acid (ortho- CAS #579-75-9, meta- CAS #586-38-9, para- CAS #100-09-4), salicylic acid (CAS #69-72-7), benzoic acid (CAS # 65-85-0), 5-nitroanthranilic acid, anthranilamide, benzamide, o- cabroethoxybenzene sulfonamide, isatoic anhydride and 4- ketobenztriazine were obtained from Aldrich Chemical Co, Milwaukee, Wisconsin.

The procedure of Example 2 was followed. .Results.—Of all the single substitutions of electron donating groups for benzoic acid derivatives, only the amino isomeric substitutions showed repellent effects. Consumption for all concentrations of benzoic acid, salicylic acid, and isomers of methoxy benzoic acid were all similar to pretreatment consumption levels (Table 2, Figures 6a - 6b & 7a - 7c, P>0.05). Consumption of water decreased as a function of increasing concentration for meta- and 4- aminobenzoic acid (Figures 8a - 8c, F=5.55, P = 0.004 and F= 3.59, P = 0.021, df=5,17, respectively) . Anthranilic acid was the most effective repellent of the benzoic acid series (F= 24.29, P < 0.001, df = 5,17). Post-hoc tests indicated consumption levels were similar for 0.5, 0.25 and 0.13% concentration and that consumption for this group differed from the control and 0.03 and 0.06 % concentrations. Most striking was the magnitude of drinking suppression, nearing zero consumption for the highest concentrations tested. Table 2

Differences in water consumption among concentrations tested

Compound F^a P post-hoc Scheffe test^c

OAA 24.29 .001 .5 0.25 .13 .06 .03 0.00

.001 .5 0.00 .06 .25 .03 0.13

4ABA 0.97

SA 2.52 BA 1.59

° F value from a one-way anova.

P probability associated with the anova that consumption across all concentration groups was equal. Rejection of the hypotheses occurred when P<0.05. ^c Post-hoc test of significant differences (P<0.05) among concentration groups (values listed in Table) . Similarities for group consumption for concentration tested are connected by lines.

Consumption did not differ from control levels for isatoic anhydride, o-carboethoxybenzene sulfonamide, 2- aminobenzamide, and 5-nitroanthranilic acid within the heterocyclic series (Table 3, Figures 9a - 9c & 10a -10c, P>0.05). Consumption was not affected by isatoic anhydride or o-carboethoxybenzene sulfonamide (P>0.05). Only consumption for 4-ketobenztriazine showed any repellent effects (Figures 10a -10c, F=8.19, df= 5,30, P < 0.001). Only the highest concentration did not significantly differ from zero consumption, though reduced consumption was evident for concentrations as low as 0.05%. Table 3 Differences in water consumption among concentrations tested

Compound F^a P^b post-hoc Scheffe test^c

5NAA 0.69 .691 .500 .005 .01 .050 .100 .001

F value from a one-way anova.

P probability associated with the anova that consumption across all concentration groups was equal. Rejection of the hypotheses occurred when P<0.05.

Post-hoc test of significant differences (P<0.05) among concentration groups (values listed in Table) . Similarities for group consumption for concentration tested are connected by lines.

Example 4. Feeding Trials

Birds.—Twenty-fouradultmale European starlingswere tested with each aminoacetophenone isomer. This bird species was chosen for use (a) because they show good chemical sensing ability (Mason, J.R. , et al., supra , (1983); Clark, L. , supra , (1987) (b) because comparable data exist concerning the responses of starlings to other avian repellents (anthranilate derivatives; Mason, J.R. , et al., "Field trial of dimethyl anthranilate as a nontoxic bird repellent in cattle and swine feedlots", J. Wildl . Manage . 49:636-642 (1985); Mason, J.R. , et al., supra, (1989); Glahn, J.F., supra , (1989) and mammalian irritants (e.g., capsaicin, zingerone, gingerol, allyl isothiocyanate; Mason, J.R. , et al., "Chemical nociception in birds". Pages 309-322 in B. Green and J.R. Mason, eds. Chemical nociception in the nose and mouth . Marcel Dekker, New York, N.Y., 361pp (1990), and (c) because starlings are agricultural pests. Bailey, E.P. , "Abundance and activity of starlings in winter in northern Utah", Condor 68:152-162. (1966); Baily, E.P., et al. "Costs of wintering starlings and red-winged blackbirds at feedlots", J. Wildl. Manage, 32:179-180 (1968); Besser, J.F., et al. , supra (1968). Chemicals.—OAAP (CAS # 551-93-9), para- aminoacetophenone (PAAP; CAS # 99-92-3) , meta- aminoacetophenone (MAAP; CAS # 99-03-6) , and alpha- aminoacetophenone (AAAP; CAS # 5468-37-1) were obtained from Sigma Chemical Company (St. Louis, MO.). Each was mixed with Purina Flight Bird Conditioner (PFBC; Purina Mills, ST. Louis, MO.) to produce the following series of concentrations: 1.0%, 0.5%, 0.1%, and 0.01% (g/g) . Procedur .

(a) Two-cup tests.—The procedures detailed in Mason et al., supra, (1989) for 2-cup avian repellency evaluations were followed. Briefly, for each isomer, 24 starlings were randomly selected, weighed, and then assigned to 4 groups

(n=6/group) on the basis of mass. Specifically, the heaviest bird was assigned to the first group, the next heaviest to the second group, and so on. During the 4-day pre-treatment period, all food was removed from the cages within 1 hour of light onset. Next, 2 cups, each containing 50 g of PFBC were placed in the front center of each cage. Cups were bound together with a rubber band to reduce spillage, and consumption was assessed after 2 hours. After testing, and until light onset of the following day, birds had free access to feed.

On the day following the last pre-treatment day, a 4- day treatment period began. Within 1 hour of light onset, each group was given 2 cups. One contained 50 g of PFBC thoroughly mixed with different quantities of an aminoacetophenone isomer. The other contained 50 g of plain PFBC. Cups were bound together with a rubber band, and cup positions were alternated daily. Groups 1-4 received PFBC containing 1.0%, 0.5%, 0.1% or 0.01% aminoacetophenone isomer, respectively. As in pre-treatment, consumption was measured after 2 hours. At the end of the fourth treatment trial, all birds were re- weighed to assess whether any change from pre-treatment mass had occurred.

Results were analyzed in 2 ways using parametric statistics. First, treatment consumption was assessed in a 3- factor analysis of variance (anova) with repeated measures on the second (days) and third (cups) factors. Next, preference ratios were calculated by dividing the aminoacetophenone consumption of each bird by total consumption (for pre¬ treatment, consumption from a randomly chosen cup by each bird was divided by total consumption). A ratio of 1.0 indicated complete preference, 0.5, indifference, and 0.0, complete rejection of the treated cup. Ratios were assessed in a 3- factor anova with repeated measures on the second (period) and third (days) factors. In all cases, the post-hoc tests (Winer 1962:198) were used to isolate significant differences among means (£ < 0.05) .

(b) One-cup tests.— he procedures detailed in Mason, J.R., et al., supra , (1989) for 1-cup avian repellency evaluations were followed. Briefly, for each isomer, 24 starlings were randomly selected, weighed, and then assigned to 4 groups (n=6/group) as described above. On the day following group assignment, a 4 day pre-treatment period began, identical in all respects to the 2-cup pre-treatment period, except that each bird was presented with only 1 cup containing 50 g of PFBC. A 4 day treatment period immediately followed pre-treatment, and during each treatment, each group was presented with 50 g samples of PFBC adulterated with a different amount of aminoacetophenone isomer (1.0%, 0.5%, 0.1%, 0.01% for groups 1-4, respectively). Consumption was recorded after 2 hours. Birds had free access to plain PFBC and water during the night. At the end of the fourth treatment trial, all birds were reweighed.

Results were analyzed in 2 ways using parametric statistics. First, a 3-factor anova with repeated measures on the second (periods) and third (days) factors was used to assess consumption. Next preference ratios were calculated by dividing the treatment consumption of each bird by total pre- treatment and treatment consumption. Ratios were assessed in a 2-factor anova with repeated measures on the second factor (days) . The post-hoc tests were used to isolate significant differences among means (£ < 0.05). Results

Ortho-aminoacetophenone.—Analysis of 2-cup tests failed to reveal differences among concentrations, although there were differences between cups (F = 105.6; l,20df; £ < 0.00001) . There also was an interaction between concentrations and cups (F = 3.53; 3,20df; £ < 0.03). All concentrations of OAAP reduced consumption relative to consumption of plain feed. However, differential consumption was least for birds presented with the 0.1% treatment (Figures 11a - lib). When preference ratios were assessed, there were significant differences among concentrations (F = 8.83; 3,20df; £ < 0.0009) and between periods (F = 32.46; l,20df; £ < 0.0001). Treatment preference ratios (mean = 0.21) were significantly lower than pre-treatment ratios (mean = 0.44). The main effect for concentration (collapsed across pre-treatment and treatment periods) reflected the fact that the mean ratio for 0.1% (0.45) was significantly greater than those for 1.0% (0.25), 0.5% (0.22), or 0.01 (0.38) (Figures 12a - 12b).

Analysis of 1-cup tests revealed significant differences in consumption between periods (F = 42.38; l,20df; £ < 0.00001) and an interaction between periods and days (F = 7.61; 3,60df; £ < 0.0O04). All concentrations of OAAP effectively reduced consumption during treatment. Examination of the interaction term showed that while consumption increased over days during the pre-treatment period, it was low and stable during treatment (Figures 11a - lib) . Analysis of preference ratios failed to reveal any significant differences. All OAAP concentrations produced ratios < 0.4 (Figures 11a - lib) .

Para-aminoacetophenone.—Analysis of 2-cup tests showed no differences among concentrations, although there were differences between cups (F = 90.34; l,20df; £< 0.00001) and among days (F = 9.26; 3,60df; £ < 0.0001) . Also, there was a significant interaction between these terms (F » 4.66; 3,60df; £ < 0.006). All PAAP concentrations reduced consumption relative to that of untreated feed (Figures 13a - 13b) . Examination of the days effect showed that consumption on day 2 (collapsed over concentrations and cups) was significantly lower than consumption on any other days. Post- hoc analysis of the interaction term failed to reveal any significant differences. Examination of preference ratios revealed a significant difference between periods (F - 88.38; l,20df; £ < 0.00001). All PAAP treatment concentrations produced ratios < 0.3 (mean = 0.20) whereas pre-treatment ratios hovered around 0.50 (mean = 0.53) (Figures 12a - 12b).

Analysis of 1-cup test consumption revealed significant differences between periods (F = 11.27; l,20df; £ < 0.003) and among days (F = 19.25; 3,60df; £ < 0.00001). Also, there was a significant interaction between these terms (F = 4.95; 3,60df; £ < 0.004). Overall, PAAP reduced consumption during treatment (Figures 13a -13b) . The days main effect (collapsed over concentrations and periods) reflected significantly higher consumption on days 3 and 4 than on days 1 and 2. Examination of the interaction showed that while consumption remained stable during pre-treatment, it increased during treatment, suggesting habituation to PAAP. Analysis of preference ratios failed to reveal any significant differences. All PAAP concentrations produced ratios > 0.4, suggesting weaker avoidance than had been observed for OAAP (Figures 12a - 12b) .

Meta-aminoacetophenone.—Analysis of 2-cup tests showed that there were no differences among concentrations, although there were differences between cups (F - 185.47; l,20df; £ < 0.00001) (Figures 13a - 13b) . Post-hoc examination of this effect revealed that all concentrations of MAAP reduced consumption relative to consumption of untreated feed. Examination of reference ratios uncovered significant differences between periods (F = 86.06; l,20df; £ < 0.00001). All MAAP concentrations produced ratios < 0.25 (mean ■ 0.198) whereas treatment preference ratios hovered around 0.5 (mean = 0.558) (Figures 13a -13b).

Analysis of 1-cup tests revealed a slight but significant difference between periods (F = 6.86; l,20df; £ < 0.016); less was consumed during treatment. When preference ratios were examined, there were no significant differences.

All MAAP concentrations produced ratios > 0.44 (Figures 13a -

13b) .

Alpha-aminoacetophenone.—Analysis of 2-cup consumption showed that there were differences between cups

(F = 381.88; l,20df; £ < 0.00001) and an interaction between days and cups (F = 8.23; 3,60df; £ < 0.0003). Post-hoc examination of these effects revealed that all concentrations of AAAP reduced consumption relative to consumption of untreated feed (Figures 14a - 14b) . However, this effect diminished during the course of treatment. Examination of preference ratios revealed significant differences between periods (F = 248.76; l,20df; £ < 0.00001). All MAAP concentrations produced ratios < 0.25 (mean = 0.193) whereas treatment preference ratios hovered around 0.5 (mean = 0.47)

(Figures 14a - 14b) .

Analysis of 1-cup consumption revealed significant differences among concentrations (F = 3.46; 3,20df; £ < 0.035), and periods (F = 121.63; l,20df; £ < 0.00001). Also, there were significant interactions between concentrations and periods (F = 8.8; 3,20df; -£ < 0.0009) and periods and days (F = 5.71; 3,60df; £ < 0.002). Post-hoc examination of the main effects indicated that consumption increased as AAAP concentration decreased, and that overall pre-treatment consumption was greater than overall treatment consumption. Analyses of the interaction terms showed that differences in consumption between periods decreased with decreasing AAAP concentration and that while consumption was stable during pre-treatment, it increased during treatment. Summary.—All of the aminoacetophenone isomers were avoided in 2-cup tests. However, differences among isomers emerged when only 1-cup was presented (Figure 15) . Overall, OAAP produced the most reliable avoidance, regardless of testing paradigm. While all of the isomers might have utility in some animal damage control applications, OAAP shows the greatest promise. As specific practical applications, it might be possible to incorporate low levels of OAAP into livestock feeds as a bird repellent. Alternatively, OAAP could be used to deter crop depredation, or as a repellent additive to formulated agricultural chemicals or mammalian poisons (to reduce accidental ingestion by birds) .

Example 5. Coniferyl benzoate, its analogs and corresponding alcohols

Two analogs of coniferyl benzoate, cinnamyl benzoate and 3,4-dimethoxycinnamyl benzoate, were selected to study how electron donating groups on the phenyl ring affect feeding repellency. The importar *e of benzoate ester was determined by comparing the feeding epellency of coniferyl benzoate and the two aforementioned analog esters to their corresponding alcohols. Additionally, benzoic acid was tested to determine the contribution of this moiety to feeding deterrency. The relative and absolute repellency of each compound was evaluated using choice and no-choice tests.

Chemicals.—A Bruker AM-500, 5 mm probe, NMR was used to confirm all chemical syntheses.

Coniferyl benzoate was obtained by continuous liquid/liquid extraction of benzoin Siam tears #3 (Alfred L. Wolff, Paris, France) . Briefly, benzoin Siam tears were dissolved in a solution of methanol and water (90:10) (Shinobu Kato, Shiseido Laboratories, Yokohama, Japan, pers. comm.), filtered, and decanted into a large round bottom flask. The filtered solution was extracted with pentane in a high volume Kontes (Vineland, NJ) liquid/liquid extraction system. During the extraction, the solution in the collection flask was magnetically stirred and not allowed to exceed 38'C. Extraction periods lasted 24-48 hours and could be repeated up to 3 times per batch. Upon completion of the extraction, pentane was evaporated from the collected solution, and the crude coniferyl benzoate was purified by crystallization (ether/pentane at -17°C for minimum of 24 hours [Daniel Joulain, Robertet, Grasse, France, pers. comm.]). Analysis of the crystallized product by (¹H) NMR indicated that peak assignments matched those reported for coniferyl benzoate. Jakubas , et al. , "Ruffed grouse feeding behavior and its relationship to the secondary metabolites of quaking aspen flower buds," J Chem Ecol 15:1899-1917 (1989).

Coniferyl alcohol was synthesized using Lindberg's (1980) procedure, with minor modifications. First, eugenol was acetylated with acetic anhydride (2.0 eq.) in pyridine. Eugenol acetate, thus obtained, was brominated via N- bromosuσcinimide in carbontetrachloride with barium carbonate (1.5 eq.) an acid-scavenger. The bro o derivative thus obtained was dissolved in N,N-dimethylformamide. Potassium acetate (10 eq.) was added and the mixture was kept at 90^βC for one hour. After evaporating the N,N-dimethylformamide under reduced pressure, the residue was dissolved in dichloromethane, washed with water, dried, and evaporated. The crude coniferyl diacetate was deesterified with lithium tetrahydroaluminate. The resulting product was purified by column chromatograph and crystallized to get coniferyl alcohol (72-74^βC; mp 72-73'C) Allen C.F.H., et al., "A synthesis of coniferyl alcohol and coniferyl benzoate," J Am Chem Soc 71:2683-2684 (1949).

Cinnamyl benzoate was synthesized by benzoylating cinnamyl alcohol (Aldrich Chem. Co., Milwaukee, WI) . Briefly, benzoyl chloride (5:1 mol) was added to a mixture of cinnamyl alcohol in pyridine at 0°C and left to warm to room temperature (approx. 23° overnight. When the reaction was complete, methylene chloride and water were added to the reaction mixture and the layers separated. The organic layer was washed with IN hydrochloric acid, neutralized with a saturated solution of sodium bicarbonate, washed with water, dried, and evaporated using toluene and a rotary evaporator. The residue was purified by column chromatography (silica gel, hexane: ethyl acetate [10:1] and crystallized. 3, -Dimethoxycinnamyl alcohol was synthesized from 3, 4-dimethoxycinnamic acid following the procedure of Ponpipom, et al. (1987) . 3,4-Dimethoxycinnamyl benzoate was synthesized bybenzoylating 3,4-dimethoxycinnamyl alcohol usingprocedures identical to those used for cinnamyl benzoate (above) . The product was purified by column chromatography (silica gel, hexane:ethyl acetate [2:1], and crystallized in ether and pentane at -17^βC. Melting points and [ H] NMR peak assignments of the synthesized products agreed with published data for 3,4-dimethoxycinnamyl alcohol and benzoate. See Kuroyanagi, et al., "Studies on the constituents of Baccharis σenistelloides_f" Chemical and Pharmaceutical Bulletin 33:5075-5078 (1985).

Benzoic acid used in the feeding trials was obtained from Aldrich Chem. Co., Milwaukee, WI.

Diet Preparation.—Compounds to be tested were added to the bird's test feed (5:1 mixture of Chick Starter and AVN Canary/Finch diet [Purina Mills Inc., St. Louis, MO] by dissolving the compounds in ethyl ether, mixing the ether solution with the feed, and then evaporating the ether under a hood. All diets were stored at -17^βC in closed containers until they were presented to the birds. A double blind design was followed when preparing the diets to prevent possible measurement biases when measuring feed consumption. Dietary concentrations for all compounds were equimolar with 3 concentrations of coniferyl benzoate (0.4%, 1.6%, 3.2% [w/w] . Control diets were prepared by mixing the test feed with ethyl ether and evaporating the ether as described above. Coniferyl alcohol is reportedly sensitive to light and will decompose slowly upon standing (Allen and Byers 1949, Aldrich Chem. Co., Milwaukee, WI) . Therefore, its stability in feed was tested prior to the feeding trials. Four test samples were prepared by dissolving coniferyl alcohol (4 x 50 mg) Aldrich Chem. Co., Milwaukee WI) in ether and applying it to the bird's test feed (4 x 2.4g). Following evaporating of the ether, the samples were stored under standard room temperature and light conditions for 2, 4, and 24 hr periods. The forth sample was stored in a closed container for 4 days at 17"C. Samples were extracted with ether and analyzed by TLC for changes in coniferyl alcohol content. No significant change in coniferyl alcohol concentrations occurred in the - 17*C sample or in the samples held under standard conditions for up to 24 hrs.

When phenolic compounds are consumed simultaneously (as they are when a plant is eaten) , synergistic interactions can be significant. Jung and Fahey, "Effects of phenolic monomers on rat performance and metabolism," J Nutr 113:546- 556 (1983); Lindroth, et al., "Chemical ecology of the tiger swallowtail: Mediation of host use by phenolic glycosides," Ecol 69:814-22 (1988). Therefore, a comparison was made between the efficacy of pure coniferyl benzoate and Siam benzoin tears (a resin from Styrax tonkinensis) as feeding repellents. Siam benzoin tears (or gum benzoin Siam) contains coniferyl benzoate, vanillin, benzoic acids, and cinnamic acids. Analysis by HPLC (see Jakubas and Gullion, supra (1990)) indicated that Siam benzoin tears #3 (Alfred L. Wolff, Paris France) contained 80% coniferyl benzoate. Cinnamyl benzoate was not found in our benzoin Siam. Benzoin Siam tears were dissolved in ether, filtered, and applied to the test feed in a manner identical to the application of other test compounds (see above) . Application levels of benzoin Siam tears were based on its coniferyl benzoate content and were matched to the test concentrations used for pure coniferyl benzoate (0.4%, 1.6%, 3.2% [w/w].

Feeding Trials.—EuropeanStarlings (Sturnusvulgaris) were funnel-trapped in rural New Jersey and Philadelphia, PA from January to March, and transported to the laboratory. This species was used because it has good chemosensory abilities (Clark, et al., "Olfactory discrimination of plant volatiles by European starling," Animal Behavior 35:227-235 (1987)), and because it is considered an agricultural pest. Glahn, et al., "Effects of dimethylanthranilate as a bird repellent livestock feed additive," Wildlife Soc Bulletin 17:313-320 (1989).

Birds were individually caged (dimensions: 61 x 36 x 41 cm) and kept in constant temperature conditions (approx. 20^βC) , under 11:13 hour light:dark cycle. During the 2 weeks before pretreatment, birds were provided free access to a 5:1 feed mixture of Chick Starter and AVN Canary/Finch diet (Purina Mills, Inc., St. Louis, MO), and oyster shell grit (United Volunteer Aviaries, Nashville, TN) . Tap water was always available.

Each chemical was evaluated in 1-cup and 2-cup tests. In every case, a 4 day pretreatment period was followed by a 4 day treatment period. During both periods, consumption during test periods was encouraged by food depriving the birds overnight.

On each pretreatment Day 1, 24 birds were randomly assigned to 6 groups (n=4/group) . Between 0900 and 1100 hours, birds received 1 cup (3 groups) or 2 cups (3 groups) , each containing 10 g of ether-treated feed. Cups were positioned in the center of the front of each cage. At the end of the 2 hours period, food cups were removed, and consumption was recorded. Birds were then given free access to maintenance feed until lights out.

During the treatment period, one 1-cup and one 2-cup group was randomly assigned to each of the 3 chemical concentrations. During the 2 hour test period, 1-cup groups were given a single 1-cup containing 10 g of treated feed. Two-cup groups were given 2 cups, 1 containing 10 g of treated feed, and the other containing 10 g of control feed (treated with ether only, as in pretreatment) . To prevent birds from associating chemical treatments with cup position (left or right) or individual cups, the position of the cups were alternated and different cups were presented each day. At the end of the test period, food cups were removed and consumption was measured. Birds were left with free access to maintenance feed until lights out. Analysis.—For 2-cup tests, the mean consumption of treated and control feed by each bird during each experiment was calculated. Means were examined in a 3-factor analysis of variance (ANOVA) with repeated measures between cups. The independent factors were chemical and concentration. In addition, preference ratios were calculated by dividing consumption of treated feed by total consumption (treatment and control) . Ratios were examined in a 2-factor (chemical, concentration) ANOVA) . For 1-cup tests, mean pretreatment and treatment consumption by each bird in each experiment was calculated. Means were examined in a 3-factor ANOVA with repeated measures between periods. The independent factors in this analysis were chemical concentration. In all cases, Tukey Hostely Significant Difference

(HSD) tests were used to isolate significant (£<0.05) differences among means.

(a) Two-cup tests. Overall, birds ate more control (3.34+0.44 g) than treated feed (1.25±0.62) (F=244.8; 1, 62 df; £ < 0.00001) .

There were also differences among chemicals. Post-hoc examination of the interaction between chemicals and periods

(F = 6.4; 6,62 df; £ < 0.00001) showed that birds given coniferyl alcohol, 3,4-dimethoxycinnamyl alcohol, or 3,4- dimethoxycinnamyl benzoate ate less treated feed (0.6+0.3,

0.7+0.2, 0.9+0.3g) , respectively) than did birds given cinnamyl alcohol, cinnamyl benzoate, or coniferyl benzoate

(1.3+0.2, 1.4+0.2, 1.4+0.2g, respectively). However, birds given benzoic acid ate more treated feed (2.1+10.3g) than any other group (Figures 18a -18g) . Consumption of control feed among groups was inverse to the consumption of treated feed.

Finally, post-hoc examination of the interaction among chemicals, concentrations, and periods (F = 2.03; 12,62 df;

£ < 0.03) revealed the following pattern of significant effects. For benzoic acid, there were no significant differences between consumption of treated and control feeds at any but the highest concentration. For cinnamyl benzoate, there was no difference in consumption at the lowest chemical concentration, but significant differences at medium and high concentrations, for cinnamyl alcohol, coniferyl benzoate, and 3,4-dimethoxycinnamyl benzoate, differences in consumption between treated and control feed increased with increasing concentrations. For 3,4-dimethoxycinnamyl alcohol, therewere no differences in consumption at the lowest concentration, but consumption of treated feed was essentially eliminated at the medium and high concentration. For coniferyl alcohol, consumption of treated feed was strongly suppressed at all concentrations.

Discussion—Analysis of preference ratios gave a pattern of results similar to those described above (Figure 19) . First, there were significant differences among chemical (F = 9.7; 6,62 df; £ < 0.00001). Post-hoc tests showed that the lowest preference ratios were associated with coniferyl alcohol (0.13), 3,4-dimethoxycinnamyl alcohol (0.19), and 3,4- dimethoxycinnamyl benzoate (0.20). Intermediate ratios were associated with cinnamyl alcohol (0.27) and cinnamyl benzoate (0.32). The highest mean ratio was associated with benzoic acid (0.44). Second, there was inverse relationship between concentration and preference ratio size (F = 2.1; 12,62 df; £ < 0.03) Figure 19). Post-hoc tests indicated that preference ratios for all coniferyl alcohol concentrations were very low. Preference ratios for the lowest concentration of 3,4-dimethoxycinnamyl alcohol or 3,4-dimethoxycinnamyl benzoate were higher than ratios for intermediate and high concentrations. Preference ratios for the lowest concentration of cinnamyl alcohol, cinnamyl benzoate, or coniferyl benzoate were higher than ratios for intermediate for high concentrations. However, the intermediate land high concentration ratios for these compounds were greater than ratios associated with intermediate or high concentrations of coniferyl alcohol, 3,4-dimehtoxycinnamyl alcohol, or 3,4- dimethoxycinnamyl alcohol, or 3,4-dimethoxycinnamyl benzoate. Finally, preference ratios for any concentration of benzoic acid were high, and approached indifference (0.50). (b) - One-cup tests. There were significant differences between periods and among concentrations. Not surprisingly, birds ate more during pretreatment than during treatment (F = 402.4; 1,71 df; £ < 0.00001), and showed greater consumption at low concentrations than at intermediate or high concentrations ((F = 29.7; 2,71 df; £ < 0.00001).

There were significant differences among chemical ((F = 13.4;

7,71 df; £ < 0.00001), and post-hoc tests showed that overall

(pretreatment and treatment) consumption was lowest for coniferyl alcohol (2.9+0.3g) and 3,4-dimethoxycinnamyl alcohol

(3.0+0.2g). The next lowest consumption was exhibited by birds given 3,4-dimethoxycinnamyl benzoate (3.4+0.2g) or coniferyl benzoate (3.5+0.3g) . Birds given cinnamyl benzoate or cinnamyl alcohol showed still higher consumption (3.9+0.30,3.9+0. g, respectively) . However, the highest overall consumption was exhibited by birds given either benzoin Siam (4.5+0.3) or benzoic acid (4.6+0.4g).

Discussion—Post-hoc evaluation of the interaction between chemicals and periods (F = 23.6; 7,71 df; £ < 0.00001) showed that all chemical except benzoic acid significantly reduced consumption during the treatment period. Examination of the interaction between concentrations and periods ((£ = 69.7; 2,71 df; £ < 0.00001), indicated that differences in consumption between periods became greater as concentrations increased. Finally, an assessment of the interaction among chemicals, concentrations and periods ((F = 5.0; 14,71 df; £ < 0.00001) , revealed a complex pattern (Figures 20a - 20h) . At the lowest concentration, consumption differences between pretreatment and treatment periods were greatest for 3,4- dimethoxycinnamyl benzoate > cinnamyl alcohol land cinnamyl benzoate > coniferyl alcohol and coniferyl benzoate > Siam > benzoic acid. 3,4-Dimethoxycinnamyl alcohol produced no drop in consumption at the lowest concentration; rather, consumption of treated feed was slightly higher than consumption of untreated feed. At the intermediate concentration, 3,4-dimethoxycinnamyl benzoate > 3,4- dimethoxycinnamyl alcohol and benzoin Siam> cinnamyl benzoate > cinnamyl alcohol > coniferyl alcohol > coniferyl benzoate

> benzoic acid. At high concentration, benzoin Siam and cinnamyl benzoate > 3,4-dimethoxycinnamyl benzoate ? benzoic BARBARA: I DID NOT PUT THE QUESTION MARK IN. IS IT SUPPOSED TO BE THERE?? acid and 3,4-dimethoxycinnamyl alcohol > cinnamyl alcohol, coniferyl benzoate, and coniferyl alcohol.

A comparison of the preference ratios for benzoin Siam tears and coniferyl benzoate indicate that benzoin Siam tears were significantly (F = 23.04; 1,17 df; £ < 0.00003), more repellent than pure coniferyl benzoate.

Claims

Claims:

1. A method of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising applying to said material a compound, wherein said compound is alpha-aminoacetophenone, ortho-aminoacetophenone, meta-aminoacetophenone, para- aminoacetophenone, ortho-hydroxyacetophenone, ortho- methoxyacetophenone, meta-methoxyacetophenone, para- methoxyacetophenone, anthranilic acid, 3-aminobenzoic acid, 4-aminobenzoic acid, 4-ketobenztriazine, coniferyl benzoate, cinnamyl benzoate, 3-4 dimethoxycinnamyl benzoate, coniferyl alcohol, cinnamyl alcohol, or 3-4 dimethoxycinnamyl alcohol, in an amount effective to reduce the consumption or utilization of said material by said birds by a statistically significant amount.

2. The method according to claim 1 wherein the compound is an aminoacetophenone.

3. The method according to claim 2 wherein the aminoacetophenone is ortho-aminoacetophenone.

4. The method according to claim 1 wherein the compound is anthranilic acid.

5. The method according to claim 1 wherein the compound is 4-ketobenztriazine.

6. The method according to claim 1 wherein the compound is coniferyl benzoate.

7. The method according to claim 1 wherein the compound is coniferyl alcohol.

8. The method according to claim 1 wherein the compound is at least partially homogeneously dispersed in a vehicle or carrier suitable for applying said compound to said material.

9. The method according to claim 8 wherein the vehicle is a starch, oil or polymer which encapsulates the compound.

10. The method according to claim 8 wherein the carrier contains the compound in an emulsified or other form capable of being at least partially dispersed in a liquid.

11. The method according to claim 1 wherein the material is a liquid.

12. The method according to claim 11 wherein the liquid is water.

13. The method according to claim 11 wherein the liquid is non-potable water.

14. The method according to claim 11 wherein the liquid is free standing on artificial surfaces.

15. The method according to claim 14 wherein the artificial surfaces are selected from the group consisting of airport runways and parking lots.

16. The method according to claim 13 wherein the non- potable water is waste water selected from the group consisting of industrial waste water and agricultural waste water.

17. The method according to claim 16 wherein the waste water is from a mine tailing ponds.

18. The method according to claim 1 wherein the material is a bird edible.

19. The method according to claim 1 wherein the material is seed.

20. The method according to claim 1 wherein the material is an agrichemical.

21. The method according to claim 1 wherein the material is a turf or grass product.

22. The method according to claim 1 wherein the material is a crop.

23. The method according to claim 1 wherein the effective amount of the compound is that amount sufficient to reduce the consumption or utilization of said material by said birds to at least about 50 percent over the amount of said material which would otherwise be consumed or utilized by said birds without said compound applied.

24. The method according to claim 1 wherein the effective amount of the compound is that amount sufficient to reduce the consumption or utilization of said material by said birds to at least about 90 percent over the amount of said material which would otherwise be consumed or utilized by said birds without said compound applied.

25. A method for repelling birds from consuming or utilizing a non-potable water comprising applying a compound which is an ester of anthranilic acid or mixtures thereof to said non-potable water in an amount effective to reduce the consumption or utilization of said water by a statistically significant amount.

26. The method according to claim 25 wherein said ester of anthranilic acid is methyl anthranilate.

27. The method according to claim 25 wherein the non- potable water is waste water selected from the group consisting of industrial waste water or agricultural waste water.

28. The method according to claim 25 wherein the non- potable water is from mine tailing ponds.

29. The method according to claim 25 wherein the non- potable water is free standing water on artificial surfaces.

30. The method according to claim 29 wherein the artificial surfaces are selected from the group consisting of airport runways and parking lots.

31. The method according to claim 25 wherein the compound is at least partially dispersed in a vehicle or carrier suitable for applying said compound to said non- potable water.

32. The method according to claim 31 wherein the vehicle or carrier is a starch, oil or polymer which encapsulates, emulsifies or substantially uniformly disperses said compound to be applied to said non-potable water.

33. The method according to claim 25 wherein the effective amount of the compound is that amount sufficient to reduce the consumption or utilization of said water by said birds to at least about 50 percent over the amount of said water which would otherwise be consumed or utilized by said birds without said compound applied.

34. The method according to claim 25 wherein the effective amount of the compound is that amount sufficient to reduce the consumption or utilization of said water by said birds to at least about 90 percent over the amount of said water which would otherwise be consumed or utilized by said birds without said compound applied.

35. A method of identifying compounds capable of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising the steps of:

(a) selecting a compound, of unknown capability of repelling birds from consuming or utilizing said material, having one of the following core structures;

wherein; R- or R,' or R.," is an electron donating group and R₂ is an electron withdrawing group or a neutral group which group does not substantially hinder electron donation to the phenyl ring by R,; and

(b) testing said compound to confirm the capability of said compound to repel birds from consuming or utilizing said material.

36. The method of claim 35 further comprising selecting a compound in step (a) wherein R₂ is an electron withdrawing group.

37. The method of claim 32 further comprising selecting a compound in step (a) wherein R₂ is group capable of forming a hydrogen bonded ring structure with an electron donating group.

38. The method of claim 35 wherein the electron donating groups, R₁ or R.,' or R.,", contribute electrons to the phenyl ring.

39. The method of claim 35 wherein the electron donating group R, contributes electrons to the phenyl ring in the positions 2, 3 or 4 with respect to R₂.

40. The method of claim 35 wherein the electron donating group is basic.

41. The method of claim 35 wherein the substituents on R₂ are of the type that do not substantially prevent R, from donating electrons to the phenyl ring.

42. The method of claim 35 wherein there is intramolecular hydrogen bonding between R₁ and R₂ when R, is in position 2 relative to R₂.

43. The method of claim 35 wherein R, and R₂ comprise a heterocyclic ring attached to the phenyl ring.

44. The method of claim 35 wherein R₂ is a lower acyl, nitro, carboxylic acid or ester.

45. The method of claim 35 wherein the electron donating group is an amine, o-lower alkyl, N-lower alkyl, or N-di lower alkyl.

46. The method of claim 35 wherein the testing step comprises comparing the amount of material consumed or utilized by a bird with the amount of material consumed or utilized by a bird comprising an avian repellant amount of said compound in a one choice test under controlled conditions whereby a statistically significant decrease in consumption of the material with said compound confirms said compounds ability to repel birds from consuming or utilizing materials otherwise susceptible to consumption or utilization by birds.

47. A method of identifying compounds capable of repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising the steps of selecting a compound, of unknown capability of repelling birds from consuming or utilizing said material, having one of the following core structures;

wherein; R₁ or R.,' or R.," is an electron donating group and R₂ is an electron withdrawing group or a neutral group which group does not substantially hinder electron donation to the phenyl ring by R,,.

48. The method of claim 47 further comprising selecting a compound wherein R₂ is an electron withdrawing group.

49. The method of claim 47 further comprising selecting a compound wherein R₂ is group capable of forming a hydrogen bonded ring structure with an electron donating group.

50. The method of claim 47 wherein the electron donating groups, , or R^ or R,", contribute electrons to the phenyl ring.

51. The method of claim 47 wherein the electron donating group R₁ contributes electrons to the phenyl ring in the positions 2, 3 or 4 with respect to R₂.

52. The method of claim 47 wherein the electron donating group is basic.

53. The method of claim 47 wherein the substituents on R₂ are of the type that do not substantially prevent , from donating electrons to the phenyl ring.

54. The method of claim 47 wherein there is intramolecular hydrogen bonding between R., and R₂ when R,, is in position 2 relative to R₂.

55. The method of claim 47 wherein R., and R₂ comprise a heterocyclic ring attached to the phenyl ring.

56. The method of claim 47 wherein R₂ is a lower acyl, nitro, carboxylic acid or ester.

57. The method of claim 47 wherein the electron donating group is an amine, o-lower alkyl, N-lower alkyl, or N-di lower alkyl.

58. A method for repelling birds from consuming or utilizing a material otherwise susceptible to consumption or utilization by birds comprising applying a compound identified from the method of claim 32 to said material in an amount effective to reduce the consumption or utilization of said material by a statistically significant amount.