WO2013003946A1 - Compositions and methods for attracting and stimulating feeding by mice and rats - Google Patents

Compositions and methods for attracting and stimulating feeding by mice and rats Download PDF

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Publication number
WO2013003946A1
WO2013003946A1 PCT/CA2012/000638 CA2012000638W WO2013003946A1 WO 2013003946 A1 WO2013003946 A1 WO 2013003946A1 CA 2012000638 W CA2012000638 W CA 2012000638W WO 2013003946 A1 WO2013003946 A1 WO 2013003946A1
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acid
mice
composition
rats
methyl
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PCT/CA2012/000638
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French (fr)
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Stephen J. Takacs
Regine M. Gries
Gerhard G. Gries
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Contech Enterprises Inc.
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Publication of WO2013003946A1 publication Critical patent/WO2013003946A1/en

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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N31/00Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
    • A01N31/02Acyclic compounds
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/002Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/002Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
    • A01N25/004Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits rodenticidal
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N35/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
    • A01N35/02Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N35/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
    • A01N35/06Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing keto or thioketo groups as part of a ring, e.g. cyclohexanone, quinone; Derivatives thereof, e.g. ketals
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/02Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/06Unsaturated carboxylic acids or thio analogues thereof; Derivatives thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/36Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
    • A01N43/04Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
    • A01N43/06Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
    • A01N43/08Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings with oxygen as the ring hetero atom
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N61/00Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/10Animals; Substances produced thereby or obtained therefrom

Definitions

  • This invention relates to compositions and methods for attracting and stimulating feeding by mice and rats.
  • mice and rats are more numerous than humans in cosmopolitan centers, serving as reservoirs for diseases (e.g. Bubonic plague), damaging crops and consuming stored foodstuffs.
  • Attractive baits are commonly used to attract mice and rats to traps and to food laced with rodenticides, either toxicants or pharmaceuticals, such as
  • rodents Trapping is not always effective, and rodents quickly learn to avoid rodenticides by linking the breath odorants of dying conspecifics with those of their toxic meal.
  • Commonly used food baits for rodents include peanuts or peanut butter (Naganuma et al. 1973; Prakash et al. 1980; Sullivan & Sullivan 1980; Gould et al. 2007), hazelnuts (Cagnin 1985; Gigirey & Rey 1998; Berry & Alleva 2010), cheese (Kadhim et al. 1989; Weihong et al. 1999; Humphries et al. 2000; Berry & Alleva 2010), corn (Chappellier 1931, Rowe et al.
  • Peanut butter is often considered to be the best rodent bait. However, its use has been curtailed because it contains allergens that can threaten the health of some humans and pets, and effective replacements are needed.
  • the present inventors acquired food sources highly attractive to mice and rats, including various types of candy, cheese, and nuts. Headspace volatiles were collected and potentially attractive components therein were identified. In selecting candidate semiochemicals in these very complex volatile mixtures for formulation in baits, the present inventors were guided by ( ) antennal responses to compounds by cockroaches as substitute generalist omnivores, (n) the occurrence of compounds in several foodstuffs, and ( ) the olfactory sensation of our own noses, assuming that the noses of humans and rodents can perceive the same compounds.
  • mice and rats 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone) from candy; 4-hydroxy-2,5-dimethylfuran-3-one (strawberry furanone) from strawberries; butyric acid, 2,3-butadione, and 3-methylbutanal from Swiss cheese; and 6- methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and ⁇ -octalactone from hazelnuts.
  • 2-hydroxy-3-methylcyclopent-2-en-l-one maple lactone
  • 4-hydroxy-2,5-dimethylfuran-3-one strawberry furanone
  • butyric acid 2,3-butadione
  • 3-methylbutanal Swiss cheese
  • 6- methyl-4-heptanone 6-methyl-(E)-2-hepten-4-one, and ⁇ -octalactone from hazelnuts.
  • the present inventors then formulated synthetic blends of food semiochemicals in wet- rendered lard (Tenderflake, Maple Leaf Foods, Mississauga, Ontario), and tested the response of rats and mice to them in a two-choice (T-tube) laboratory olfactometer fitted with live traps in experimental and control chambers. Three proportional measures of attraction were used: percent of test animals choosing either chamber, percent of time they spent in either chamber, and percent captured in traps in either chamber.
  • mice (Mm musculus) walked significantly more often into chambers baited with synthetic maple lactone formulated in lard (treatment bait) compared to control chambers baited with lard alone. They also spent significantly more time in the chamber baited with maple lactone in lard and were caught significantly more often in the trap baited with maple lactone and lard. There was no significant difference in the number of mice that walked first toward chambers baited with maple lactone formulated in lard or Swiss cheese added to lard. Mice also spent similar durations in chambers baited with either stimulus and were captured in similar numbers in traps baited with either stimulus.
  • Two-component baits formed by adding strawberry furanone or cheese to maple lactone were significantly superior to maple lactone alone by all three measures (first choice, duration spent in chamber, and percent caught in traps baited with the two-component baits).
  • a three-component bait comprised of maple lactone, strawberry furanone and cheese was significantly more potent by all three measures than a two- component bait comprised of maple lactone and strawberry furanone.
  • a complex six-component bait comprised of maple lactone, two cheese volatiles (2,3- butadione and 3-methylbutanal) and three hazelnut volatiles (6-methyl-4-heptanone, 6- methyl-(E)-2hepten-4-one and ⁇ -octalactone) formulated in lard was significantly more attractive by all three measures to rats ⁇ Rattus norweigicus) than was lard alone.
  • a six-component bait comprised of maple lactone (0.01%), two cheese volatiles [2,3- butadione (0.01%) and 3-methylbutanal (0.1%)], and three hazelnut volatiles [6-methyl- 4-heptanone (0.01%), 6-methyl-(E)-2hepten-4-one (0.01%) and ⁇ -octalactone (0.01%)] formulated in lard was similarly attractive to mice (Mus musculus) and rats (Rattus norvegicus) by all three measures as baits previously tested at a 10 times greater dose. These results demonstrate that multiple-component food-volatile baits for rodents retain their potency at a range of low doses.
  • mice Male musculus
  • rats Ratitos
  • the six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl- 4-heptanone, 6-methyl-(£)-2-hepten-4-one, and ⁇ -octalactone was at least twice as attractive as Provoke®.
  • a modified (new) six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en- 1-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-(E)-2-hepten-4-one, and y- octalactone was comparable to the original six-component bait comprised of 2-hydroxy- 3-methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and ⁇ -octalactone in attracting mice (Mus musculus) in greater numbers than to control baits comprised of Provoke® Mouse Attractant.
  • the present inventors searched for a feeding stimulant to add to the synthetic chemical bait.
  • the present inventors ran many feeding trials with a wide range of carefully selected foodstuffs, including oats, honey oats, wheat, barley, rice, peas, lentils, beans, corn, starch, pudding, molasses, gelled peanut oil, baby food, pork rinds, beef tallow, dry-rendered lard, and cracklings. Dry-rendered lard and cracklings were the most preferred food types, and were selected for admixture to the new six-component bait.
  • a composition comprising the new six-component bait (50%), dry-rendered lard (40%) and cracklings (10%) was prepared and then tested to determine whether female house mice and rats preferred to feed on this composition or on a composition containing only the new six-component synthetic food bait. Both mice and rats consumed significantly more bait containing dry-rendered lard and cracklings than the new six-component bait without these constituents. These results confirmed the feeding stimulatory effect of dry- rendered lard and cracklings, and showed that greater feeding stimulation can be achieved by a composition that includes synthetic food constituents, dry-rendered lard and cracklings than can be achieved by the synthetic food composition alone.
  • the composition of the experimental bait was: the new six-component bait (1/100 dilution) (50%), dry-rendered lard (40%) and cracklings (10%).
  • FIGURE 1 illustrates the experimental design used to test behavioural responses of adult male and female rats and mice to semiochemical lures according to an embodiment of the invention.
  • Numbers refer to components of the experimental design, as follows: (1) aquarium (30 ⁇ 30 ⁇ 60 cm) for housing a test animal prior to the onset of a bioassay; (2) aluminum gate with opening mechanism; (3) T-tube (75 ⁇ 50 ⁇ 10 cm); (4a, 4b) aquaria (30 30 x 60 cm each) housing a live trap (5a, 5b) baited with test stimulus.
  • FIGURE 2 illustrates (a) the percent of mice (Mus musculus) in Experiment 1 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention [lard (100 mg) + synthetic maple lactone (1%)] or the control bait [lard (100 mg)] which were randomly assigned to, and placed in, 5a or 5b.
  • Percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test.
  • FIGURE 3 illustrates (a) the percent of mice (Mus musculus) in Experiment 2 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention [lard (100 mg) + Swiss cheese (100 mg)] or the control bait [lard (100 mg) + synthetic maple lactone (1%)] which were randomly assigned to, and placed in, 5a or 5b.
  • P > 0.05 for the treatment bait.
  • FIGURE 4 illustrates (a) the percent of mice (Mus musculus) in Experiments 3-5 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b.
  • lard was tested at 100 mg
  • synthetic maple lactone and strawberry furanone were tested at 1% each
  • Swiss cheese was tested at 100 mg.
  • FIGURE 5 illustrates (a) the percent of mice (Mus musculus) in Experiments 6-7 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b.
  • lard was tested at 100 mg
  • synthetic maple lactone was tested at 1%
  • synthetic cheese volatiles (CV) were tested at 0.3 % each.
  • Ground-up hazelnut was tested at 100 mg.
  • FIGURE 6 illustrates (a) the percent of mice (Mus musculus) in Experiments 8-9 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b.
  • lard was tested at 100 mg
  • synthetic maple lactone was tested at 1%
  • the two synthetic cheese volatiles (CV) were tested at 0.3% each
  • HV three synthetic hazelnut volatiles
  • FIGURE 7 illustrates (a) the percent of rats (Rattus norvegicus) in Experiment 10 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the time rats spent in 4a or 4b, and (c) the percent of rats that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b.
  • lard was tested at 100 mg and synthetic maple lactone at 1%.
  • the two synthetic cheese volatiles (CV) were tested at 0.3% each, and the three synthetic hazelnut volatiles (HV) were tested at 0.1% each.
  • FIGURE 8 illustrates (a) the percent of mice (Mus muscalus) in Experiment 11, and the percent of rats ⁇ Rattus norweigicus) in Experiment 12, entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice or rats spent in 4a or 4b, and (c) the percent of mice or rats that entered the live trap 5a or 5b (see FIGURE 1) in response to a six-component bait according to an embodiment of the invention or control (lard) bait which was randomly assigned to, and placed in, 5a or 5b.
  • Percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three pairs of bars in each of
  • FIGURE 9 illustrates (a) the response of mice (Mus muscalus) in Experiment 13, and the response of rats (Rattus norvegicus) in Experiment 14, entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice or rats spent in 4a or 4b, and (c) the percent of mice and rats that entered the live trap 5a or 5b (see FIGURE 1) in response to the six-component bait according to an embodiment of the invention or the control
  • FIGURE 10 illustrates (a) the response of mice (Mus muscalus) in Experiments 15 and 16 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to a modified (new) six-component bait (Experiment 15) or the original six-component bait according to an embodiment of the invention (Experiment 16) in comparison to the control (Provoke Mouse) bait which was randomly assigned to, and placed in, 5a or 5b.
  • FIGURE 11 illustrates (a) the percent of mice (Mus musculus) in Experiment 17 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered live trap 5a or 5b (see FIGURE 1) in response to either the bait composition of a new six-component bait plus the two- component blend of the body odorants lactic acid and oleic acid (treatment stimulus) according to an embodiment of the invention or in response to Provoke® Mouse
  • Attractant control stimulus which were randomly assigned to, and placed in, 5a or 5b.
  • Percent first-choice and percent trapped data were analyzed by ⁇ test with Yates correction, whereas percent time-spent data were analyzed by Student's t-test.
  • bars with an asterisk (*) indicate a statistically significant preference (P ⁇ 0.05) for the treatment stimulus.
  • FIGURE 12 illustrates the feeding preference of mice (Mus musculus) in Experiment 18, and of rats (Rattus norvegicus) in Experiment 19, when presented with a choice between a new six-component bait according to an embodiment of the invention [maple lactone (0.01%) butyric acid (0.01%), ⁇ -octalactone (0.01%), 2,3-butadione (0.01%), 3- methylbutanal (0.1%), 6-methyl-(E)-2-hepten-4-one (0.01%)] formulated in wet-rendered lard with or without dry-rendered lard and cracklings. Both mice and rats consumed significantly more bait with dry-rendered lard and cracklings (t-test; P ⁇ 0.05), as indicted by the asterisks (*) on bars.
  • FIGURE 13 illustrates the percent of female mice (Mus musculus) in Experiment 20, and the percent of male mice in Experiment 21, that walked first toward and fed first on the bait composition with dry-rendered lard and cracklings (treatment bait according to an embodiment of the invention) or Provoke (control bait).
  • the composition of the new six- component bait was identical to that described in the caption of FIGURE 12 except that it was diluted by 1/100.
  • Percent first-choice and percent first-feeding data were analyzed by ⁇ test with Yates correction. In Experiment 20, bars with an asterisk (*) indicate a significant preference for the treatment bait (P ⁇ 0.05).
  • Lard is the melted and clarified fat of pigs.
  • leaf lard made from the fatty deposit surrounding the kidneys of a pig
  • fatback lard made from the subcutaneous fat on the back of a pig
  • soft caul lard made from fat surrounding the digestive organs.
  • lard can be made from any fatty tissue of a pig, e.g. cheeks cut from
  • Lard can be wet-rendered by boiling fatty tissue, or dry-rendered by heating fatty tissue in the absence of water.
  • wet-rendered lard is referred to as lard, unless clarification is needed to ensure that it is distinguished from dry-rendered lard. Cracklings are the crisp residue left after lard has been dry-rendered.
  • mice and rats were selected: maple lactone (2-hydroxy-3-methylcyclopent-2-en-l-one) from candy; strawberry furanone (4-hydroxy-2,5-dimethylfuran-3-one) from strawberries; butyric acid, 2,3-butadione, and 3-methylbutanal from Swiss cheese; and 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one and ⁇ -octalactone from hazelnuts.
  • the present inventors tested behavioral responses of ( ) >12-week-old reproductively active adult male and female rats (Rattus norvegicus), and (/ ) >8-week-old
  • T-tube olfactometer (FIGURE 1).
  • the experimental set-up consisted of 3 glass aquaria (30 x 30 * 60 cm each) interconnected by a T-tube 3 (75 ⁇ 50 cm; 10 cm diam). It was housed in a beige enclosure (3 2 1.3 m; not shown) with a small observation port. For each replicate, a single rodent was deprived of food (but not water) for 8 h, and then placed into aquarium 1 which was illuminated by a 20-W red bulb to facilitate observations of the animal's position. Following 1 h of acclimation aluminum gate 2 was opened and the animal allowed to enter the stem of T-tube 3 in response to semiochemical baits randomly assigned to live trap 5a or 5b in the outer corner of aquaria 4a or 4b.
  • mice Attraction of mice to two- or three-component baits
  • Experiments 3-5 explored potential additive or synergistic effects of several components formulated in lard on the attraction of mice.
  • Experiments 3 and 4 tested whether two-component baits are more effective in attracting mice than maple lactone as a single-component bait.
  • Experiment 3 compared the response of mice to synthetic 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone) alone or with synthetic 4-hydroxy-2,5-dimethylfuran-3-one (strawberry furanone), and
  • Experiment 4 compared the response of mice to synthetic maple lactone alone or with cheese.
  • mice unexpectedly walked significantly more often first toward the two-component baits, spent significantly more time near them, and entered significantly more often the traps baited with them (FIGURE 4).
  • Experiment 5 Compared the response to a three- component bait (synthetic maple lactone plus synthetic strawberry furanone plus cheese) with that to a two-component bait (synthetic maple lactone plus synthetic strawberry furanone).
  • mice are significantly more attracted to complex multi- component food odors than to simple ones.
  • mice Attraction of mice to three-, four- or five-component baits Taking into account that a three-component bait in Experiment 5 was more attractive to mice than a two-component bait, Experiment 6 tested whether an even more complex five-component bait, comprising synthetic 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone), three synthetic cheese volatiles (CV) (butyric acid, 2,3-butadione, and 3- methylbutanal) and ground-up hazelnut, was more attractive to mice than the same bait lacking hazelnut. In Experiment 6, mice walked significantly more often first toward the five-component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 5).
  • mice are strongly attracted to complex odorants, particularly those that include volatile components of hazelnuts or other nuts that contain similar components.
  • the deletion of butyric acid without diminishing the degree of attraction suggests that some degree of redundancy occurs among the components of complex odorants.
  • hazelnut as part of a complex bait greatly contributed to the attraction of mice, but non-volatile components are allergenic to some people and pets.
  • Experiments 8 and 9 tested whether hazelnut could be replaced by two or three non-allergenic key hazelnut volatiles (HV) (6-methyl-4-heptanone; 6-methyl-(E)-2- hepten-4-one; ⁇ -octalactone) without diminishing the synergistic effect of hazelnut.
  • HV non-allergenic key hazelnut volatiles
  • mice walked more often first toward the bait containing two (Experiment 8) or three (Experiment 9) synthetic hazelnut volatiles, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 6).
  • mice are highly responsive to volatiles that indicate the presence of nutritious hazelnuts, as well as other nuts that contain similar volatile constituents.
  • Experiment 10 tested the response of rats ⁇ Rattus norvegicus) to the same six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en- 1 -one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and ⁇ -octalactone.
  • rats unexpectedly walked significantly more often first toward the six- component bait formulated in lard than to lard alone (FIGURE 7). They also spent significantly more time near the six-component bait, and entered significantly more often the trap baited with it (FIGURE 7).
  • the dose of odorants emanating from baits affects both the space over which animals are attracted and the proportion of them entering a trap. A higher dose may be more effective in long-range attraction of animals but may become too overwhelming to mediate short-range orientation towards, and entry into, a trap.
  • a ten-fold lower dose of the six-component bait comprised of 2-hydroxy-3- methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6- methyl-(E)-2-hepten-4-one, and ⁇ -octalactone was tested to see if it would retain its effectiveness against mice (Experiment 11) and rats (Experiment 12).
  • mice and rats walked more often first toward the low-dose six- component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 8). Thus the dose could be lowered by without affecting the positive response of both mice and rats.
  • mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus musculus) against mice (Mus
  • mice in Experiment 13 walked more often first toward the bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 9).
  • 6-methyl-4-heptanone was removed and replaced with butyric acid to form a modified (new) six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, butyric acid, 2,3-butadione, 3- methylbutanal, 6-methyl-(£)-2-hepten-4-one, and ⁇ -octalactone.
  • said new six-component bait was tested with mice ⁇ Mus musculus) against Provoke® Mouse Attractant, and for comparison in Experiment 16, the original six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, 2,3- butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and ⁇ - octalactone was also tested with mice ⁇ Mus musculus) against Provoke® Mouse
  • Body swabs were taken from lactating females and infants of both house mice (Mus musculus) and Norway rats (Rattus norvegicus) and analysed by coupled gas
  • Experiment 17 was done to determine the effect of these body odor components on enhancing the attractiveness of the new six-component blend (see EXAMPLE 11).
  • Two representative components, lactic acid and oleic acid, were selected and formulated at a 1 :9 ratio in wet-rendered lard at 1% in combination with the new six-component bait.
  • mice walked significantly more often first toward the new six- component bait with oleic and lactic acid (pheromone constituents) added than they did to the new six-component bait alone (FIGURE 11). They also spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 11), clearly indicating that body odorants enhance bait effectiveness.
  • Experiment 17 further support our findings that rodents are attracted by complex chemical stimuli, and demonstrate that combining body odor pheromone with a food bait will enhance attraction to a level greater than that achieved by a food bait alone.
  • Experiment 20 tested a reduced dose (1/100 dilution) of the new six-component bait in combination with dry-rendered lard and cracklings, with Provoke® Mouse Attractant as a control stimulus.
  • the composition of the experimental bait was: the new six-component blend (1/100 dilution) (50%), dry-rendered lard (40%) and cracklings (10%).
  • the experiment was run in the T-tube olfactometer (see EXAMPLE 2, FIGURE 1).

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Abstract

This invention pertains to novel compositions of compounds derived from sugar-containing confections, strawberries, cheese, nuts and lactating female mice and rats. Said compositions can be used alone or in combination with dry-rendered lard and cracklings to attract and induce feeding by mice and rats. A novel composition comprising 2-hydroxy-3-methylcyclopent-2-en-1-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-(E)-2-hepten-4-one, γ-octalactone, dry-rendered lard and cracklings produced during the dry rendering process was superior to a leading commercial bait in attracting and inducing feeding by mice and rats. Said novel compositions can be used in traps designed to capture mice or rats, and can also be used in foodstuffs laced with a rodenticide.

Description

COMPOSITIONS AND METHODS FOR ATTRACTING AND STIMULATING FEEDING BY MICE AND RATS
CROSS-REFERENCE TO RELATED APPLICATION
This application claims the benefit of U.S. provisional application No. 61/504,280, filed 4 July 2011, which is incorporated herein by reference.
FIELD OF THE INVENTION
This invention relates to compositions and methods for attracting and stimulating feeding by mice and rats.
BACKGROUND OF THE INVENTION
Mice and rats ("rodents") are more numerous than humans in cosmopolitan centers, serving as reservoirs for diseases (e.g. Bubonic plague), damaging crops and consuming stored foodstuffs. Attractive baits are commonly used to attract mice and rats to traps and to food laced with rodenticides, either toxicants or pharmaceuticals, such as
chemosterilants. Trapping is not always effective, and rodents quickly learn to avoid rodenticides by linking the breath odorants of dying conspecifics with those of their toxic meal. Commonly used food baits for rodents include peanuts or peanut butter (Naganuma et al. 1973; Prakash et al. 1980; Sullivan & Sullivan 1980; Gould et al. 2007), hazelnuts (Cagnin 1985; Gigirey & Rey 1998; Berry & Alleva 2010), cheese (Kadhim et al. 1989; Weihong et al. 1999; Humphries et al. 2000; Berry & Alleva 2010), corn (Chappellier 1931, Rowe et al. 1974; Prakash et al. 1980; Baumler & Asran 1987; Wang & Wang 1997, 1999; Chopra et al. 2009), wheat (Schander & Gotze 1930; Chappellier 1931 ; Naganuma et al. 1973; Rowe et al. 1974; Mainardi et al. 1975a, b; Sidiqui & Khan 1982; Ahman & Parshad 1985; Baumler & Asran 1987; Shafi et al. 1992; Wang & Wang 1997, 1999; Chopra et al. 2009), rice (Chappellier 1931; Mainardi et al. 1975b; Wang & Wang 1997, 1999; Robards & Saunders 1998), oats (Chappellier 1931 ; Rowe et al. 1974;
Sullivan & Sullivan 1980; Partridge 1981 ; Klemann & Pelz 2005), barley (Chappellier 1931; Mainardi et al. 1975; Wang & Wang 1997, 1999; Rustamani et al. 2005), potato (Schander & Gotze 1930; Naganuma et al. 1973; Sarker et al. 1988; Wang & Wang 1997; Imaizumi et al. 2001), and fish (Naganuma et al. 1973; Rowe et al. 1974; Otsu 1979; Robards & Saunders 1998; Pervez 2007).
In the above-cited references, consumption of bait is often used as the indicator of bait effectiveness. This, however, is rarely indicative of bait attractiveness.
There are various US patents for rodent baits formulated with or without rodenticides. Lusby (1972) and Link (1957, 1963) teach the use of combinations of Warfarin, grains and sugars as baits. Ware (1975) concludes that oats, corn and sugar are effective baits.
Hollis (1989) reports artificial maple concentrate as a rodent attractant. Youngdale (1977) and Lazar and Lira (1986) teach the use of mixtures of chemosterilents and sugar, or rodenticides and artificial sweetener, as acceptable baits. Dreikorn (1979) claims rodenticidal N-pyridyl-N-phenylamines formulated in honey, molasses, corn syrup and standard animal feed for rodent control. Galef, Jr. et al. (1989) demonstrate that constituents of rodent exhalants improve bait acceptance.
Peanut butter is often considered to be the best rodent bait. However, its use has been curtailed because it contains allergens that can threaten the health of some humans and pets, and effective replacements are needed.
Improved attractive baits for effectiveness of rodent control programs are desirable. SUMMARY OF THE INVENTION
The present inventors acquired food sources highly attractive to mice and rats, including various types of candy, cheese, and nuts. Headspace volatiles were collected and potentially attractive components therein were identified. In selecting candidate semiochemicals in these very complex volatile mixtures for formulation in baits, the present inventors were guided by ( ) antennal responses to compounds by cockroaches as substitute generalist omnivores, (n) the occurrence of compounds in several foodstuffs, and ( ) the olfactory sensation of our own noses, assuming that the noses of humans and rodents can perceive the same compounds.
Taking this approach, the present inventors have identified as candidate attractive semiochemicals for mice and rats: 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone) from candy; 4-hydroxy-2,5-dimethylfuran-3-one (strawberry furanone) from strawberries; butyric acid, 2,3-butadione, and 3-methylbutanal from Swiss cheese; and 6- methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone from hazelnuts. The present inventors then formulated synthetic blends of food semiochemicals in wet- rendered lard (Tenderflake, Maple Leaf Foods, Mississauga, Ontario), and tested the response of rats and mice to them in a two-choice (T-tube) laboratory olfactometer fitted with live traps in experimental and control chambers. Three proportional measures of attraction were used: percent of test animals choosing either chamber, percent of time they spent in either chamber, and percent captured in traps in either chamber.
Mice (Mm musculus) walked significantly more often into chambers baited with synthetic maple lactone formulated in lard (treatment bait) compared to control chambers baited with lard alone. They also spent significantly more time in the chamber baited with maple lactone in lard and were caught significantly more often in the trap baited with maple lactone and lard. There was no significant difference in the number of mice that walked first toward chambers baited with maple lactone formulated in lard or Swiss cheese added to lard. Mice also spent similar durations in chambers baited with either stimulus and were captured in similar numbers in traps baited with either stimulus.
Two-component baits formed by adding strawberry furanone or cheese to maple lactone were significantly superior to maple lactone alone by all three measures (first choice, duration spent in chamber, and percent caught in traps baited with the two-component baits). In addition, a three-component bait comprised of maple lactone, strawberry furanone and cheese was significantly more potent by all three measures than a two- component bait comprised of maple lactone and strawberry furanone.
Multiple-component baits formed by adding ground up hazelnuts to baits composed of maple lactone, and three cheese volatiles (butyric acid, 2,3-butadione and 3- methylbutanal) were significantly more attractive by all three measures than the same multiple-component bait with no ground up hazelnuts. Similarly, a simpler multiple- component bait comprised of maple lactone, two cheese volatiles (2,3-butadione and 3- methylbutanal) and ground up hazelnuts was significantly more attractive to mice than the same multiple-component bait with no ground up hazelnuts.
Adding two hazelnut volatiles (6-methyl-4-heptanone and 6-methyl-(E)-2-hepten-4-one) or three hazelnut volatiles (6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one and γ- octalactone) to maple lactone and two cheese volatiles (2,3-butadione and 3- methylbutanal) caused more mice to be attracted by all three measures than simpler baits comprised of maple lactone and the two cheese volatiles. These results identified three bioactive hazelnut volatiles, and demonstrated that more complex baits are superior to less complex multiple-component baits. A complex six-component bait comprised of maple lactone, two cheese volatiles (2,3- butadione and 3-methylbutanal) and three hazelnut volatiles (6-methyl-4-heptanone, 6- methyl-(E)-2hepten-4-one and γ-octalactone) formulated in lard was significantly more attractive by all three measures to rats {Rattus norweigicus) than was lard alone.
A six-component bait comprised of maple lactone (0.01%), two cheese volatiles [2,3- butadione (0.01%) and 3-methylbutanal (0.1%)], and three hazelnut volatiles [6-methyl- 4-heptanone (0.01%), 6-methyl-(E)-2hepten-4-one (0.01%) and γ-octalactone (0.01%)] formulated in lard was similarly attractive to mice (Mus musculus) and rats (Rattus norvegicus) by all three measures as baits previously tested at a 10 times greater dose. These results demonstrate that multiple-component food-volatile baits for rodents retain their potency at a range of low doses.
The six-component bait was tested with mice (Mus musculus) against Provoke® Mouse Attractant, and with rats (Rattus norweigicus) against Provoke® Professional Rat
Attractant. For each species and by all three measures the six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl- 4-heptanone, 6-methyl-(£)-2-hepten-4-one, and γ-octalactone was at least twice as attractive as Provoke®.
A modified (new) six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en- 1-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-(E)-2-hepten-4-one, and y- octalactone was comparable to the original six-component bait comprised of 2-hydroxy- 3-methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone in attracting mice (Mus musculus) in greater numbers than to control baits comprised of Provoke® Mouse Attractant. These results reinforce the operational potential of complex odorants for rodents, and support the hypothesis that there is some redundancy among bait components. The effect of adding body odor constituents (pheromones) to a synthetic food bait composition was tested. Compounds in body swabs taken from lactating female mice and rats, as well as nursing pups were identified as: nonanoic acid (1.2%), decanoic (capric) acid (2.4%), dodecanoic (lauric) acid (17.8%), tetradecanoic (myristic) acid (5.9%), hexadecanoic (palmitric) acid (41.4%), (Z)-octadec-9-enoic (oleic) acid (17.8%), octadecanoic (stearic) acid (5.9%), lactic acid (1.8%) and glycerol (5.9%). When a 1 :9 blend of two representative pheromone constituents, lactic acid and oleic acid, respectively, was tested in the T-tube olfactometer in combination with the new six- component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, butyric acid, 2,3- butadione, 3-methylbutanal, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone, female mice walked significantly more often toward the food bait-pheromone composition than to the food bait composition alone. They also spent significantly more time near the experimental composition, and entered significantly more often the trap baited with it. These results further support our findings that rodents are attracted by complex chemical stimuli, and demonstrate that combining body odor pheromone with a food bait will enhance attraction to a level greater than that achieved by a food bait alone.
In an eleventh claim, the present inventors searched for a feeding stimulant to add to the synthetic chemical bait. The present inventors ran many feeding trials with a wide range of carefully selected foodstuffs, including oats, honey oats, wheat, barley, rice, peas, lentils, beans, corn, starch, pudding, molasses, gelled peanut oil, baby food, pork rinds, beef tallow, dry-rendered lard, and cracklings. Dry-rendered lard and cracklings were the most preferred food types, and were selected for admixture to the new six-component bait.
A composition comprising the new six-component bait (50%), dry-rendered lard (40%) and cracklings (10%) was prepared and then tested to determine whether female house mice and rats preferred to feed on this composition or on a composition containing only the new six-component synthetic food bait. Both mice and rats consumed significantly more bait containing dry-rendered lard and cracklings than the new six-component bait without these constituents. These results confirmed the feeding stimulatory effect of dry- rendered lard and cracklings, and showed that greater feeding stimulation can be achieved by a composition that includes synthetic food constituents, dry-rendered lard and cracklings than can be achieved by the synthetic food composition alone.
Adding dry-rendered lard and cracklings was tested to see if it would also enhance the attractive properties of the new six-component bait. Using the T-tube olfactometer, the present inventors tested the new six-component bait in combination with dry-rendered lard and cracklings against Provoke® Mouse Attractant as a control stimulus. The composition of the experimental bait was: the new six-component bait (1/100 dilution) (50%), dry-rendered lard (40%) and cracklings (10%). Female house mice surprisingly walked significantly more often first toward the composition bait than toward Provoke® Mouse Attractant, and also fed first significantly more often on the composition bait
Similarly, male house mice also walked more often first toward the composition bait than toward Provoke® Mouse Attractant, and fed more often on the composition bait than on Provoke® Mouse Attractant. These results show that combining a synthetic food bait (the new six-component bait) with naturally derived foodstuffs (dry-rendered lard and cracklings) enhance both attraction and feeding stimulation of rodents.
DESCRIPTION OF THE DRAWINGS In drawings which show non-limiting embodiments of the invention:
FIGURE 1 illustrates the experimental design used to test behavioural responses of adult male and female rats and mice to semiochemical lures according to an embodiment of the invention. Numbers refer to components of the experimental design, as follows: (1) aquarium (30 χ 30 χ 60 cm) for housing a test animal prior to the onset of a bioassay; (2) aluminum gate with opening mechanism; (3) T-tube (75 χ 50 χ 10 cm); (4a, 4b) aquaria (30 30 x 60 cm each) housing a live trap (5a, 5b) baited with test stimulus. FIGURE 2 illustrates (a) the percent of mice (Mus musculus) in Experiment 1 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention [lard (100 mg) + synthetic maple lactone (1%)] or the control bait [lard (100 mg)] which were randomly assigned to, and placed in, 5a or 5b. Percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three sets of paired bars, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus. FIGURE 3 illustrates (a) the percent of mice (Mus musculus) in Experiment 2 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention [lard (100 mg) + Swiss cheese (100 mg)] or the control bait [lard (100 mg) + synthetic maple lactone (1%)] which were randomly assigned to, and placed in, 5a or 5b. In each of the three sets of paired bars, there was no statistically significant preference (P > 0.05) for the treatment bait.
FIGURE 4 illustrates (a) the percent of mice (Mus musculus) in Experiments 3-5 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b. In each of Experiments 3-5, lard was tested at 100 mg, synthetic maple lactone and strawberry furanone were tested at 1% each, and Swiss cheese was tested at 100 mg. In each of Experiments 3-5, percent first-choice and percent trapped data were analyzed by a binomial test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three sets of paired bars in each of Experiments 3-5, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus.
FIGURE 5 illustrates (a) the percent of mice (Mus musculus) in Experiments 6-7 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b. In each of experiments 6-7, lard was tested at 100 mg, synthetic maple lactone was tested at 1%, and synthetic cheese volatiles (CV) were tested at 0.3 % each. Ground-up hazelnut was tested at 100 mg. In each of Experiments 6-7, percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three sets of paired bars in each of Experiments 6-7, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus. FIGURE 6 illustrates (a) the percent of mice (Mus musculus) in Experiments 8-9 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b. In each of Experiments 8-9, lard was tested at 100 mg, synthetic maple lactone was tested at 1%, the two synthetic cheese volatiles (CV) were tested at 0.3% each, and the three synthetic hazelnut volatiles (HV) were tested at 0.1% each. In each of Experiments 8-9, percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three sets of paired bars in each of Experiments 8-9, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus. FIGURE 7 illustrates (a) the percent of rats (Rattus norvegicus) in Experiment 10 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the time rats spent in 4a or 4b, and (c) the percent of rats that entered the live trap 5a or 5b (see FIGURE 1) in response to the treatment bait according to an embodiment of the invention or control bait which were randomly assigned to, and placed in, 5a or 5b. In treatment and control baits, lard was tested at 100 mg and synthetic maple lactone at 1%. The two synthetic cheese volatiles (CV) were tested at 0.3% each, and the three synthetic hazelnut volatiles (HV) were tested at 0.1% each. In each of the three sets of paired bars, bars with an asterisk (*) indicate a statistically significant preference (P< 0.05) for the treatment stimulus. Percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three sets of paired bars, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus.
FIGURE 8 illustrates (a) the percent of mice (Mus muscalus) in Experiment 11, and the percent of rats {Rattus norweigicus) in Experiment 12, entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice or rats spent in 4a or 4b, and (c) the percent of mice or rats that entered the live trap 5a or 5b (see FIGURE 1) in response to a six-component bait according to an embodiment of the invention or control (lard) bait which was randomly assigned to, and placed in, 5a or 5b. Percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed by a Students t-test. In each of the three pairs of bars in each of
Experiments 11 and 12, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus). FIGURE 9 illustrates (a) the response of mice (Mus muscalus) in Experiment 13, and the response of rats (Rattus norvegicus) in Experiment 14, entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice or rats spent in 4a or 4b, and (c) the percent of mice and rats that entered the live trap 5a or 5b (see FIGURE 1) in response to the six-component bait according to an embodiment of the invention or the control
(Provoke Mouse or Provoke Rat) bait which was randomly assigned to, and placed in, 5a or 5b. In each of Experiments 13 and 14, percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed a Students t-test. In each of the three sets of paired bars in each experiment, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus.
FIGURE 10 illustrates (a) the response of mice (Mus muscalus) in Experiments 15 and 16 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered the live trap 5a or 5b (see FIGURE 1) in response to a modified (new) six-component bait (Experiment 15) or the original six-component bait according to an embodiment of the invention (Experiment 16) in comparison to the control (Provoke Mouse) bait which was randomly assigned to, and placed in, 5a or 5b. In each of Experiments 15 and 16, percent first-choice and percent trapped data were analyzed by a binominal test, whereas percent time-spent data were analyzed a Students t-test. In each of the three sets of paired bars in each experiment, bars with an asterisk (*) indicate a statistically significant preference (P < 0.05) for the treatment stimulus. FIGURE 11 illustrates (a) the percent of mice (Mus musculus) in Experiment 17 entering first the aquarium 4a or 4b (see FIGURE 1), (b) the percent of time mice spent in 4a or 4b, and (c) the percent of mice that entered live trap 5a or 5b (see FIGURE 1) in response to either the bait composition of a new six-component bait plus the two- component blend of the body odorants lactic acid and oleic acid (treatment stimulus) according to an embodiment of the invention or in response to Provoke® Mouse
Attractant (control stimulus) which were randomly assigned to, and placed in, 5a or 5b. Percent first-choice and percent trapped data were analyzed by χ test with Yates correction, whereas percent time-spent data were analyzed by Student's t-test. In each of the three sets of paired bars, bars with an asterisk (*) indicate a statistically significant preference (P≤ 0.05) for the treatment stimulus.
FIGURE 12 illustrates the feeding preference of mice (Mus musculus) in Experiment 18, and of rats (Rattus norvegicus) in Experiment 19, when presented with a choice between a new six-component bait according to an embodiment of the invention [maple lactone (0.01%) butyric acid (0.01%), γ-octalactone (0.01%), 2,3-butadione (0.01%), 3- methylbutanal (0.1%), 6-methyl-(E)-2-hepten-4-one (0.01%)] formulated in wet-rendered lard with or without dry-rendered lard and cracklings. Both mice and rats consumed significantly more bait with dry-rendered lard and cracklings (t-test; P < 0.05), as indicted by the asterisks (*) on bars.
FIGURE 13 illustrates the percent of female mice (Mus musculus) in Experiment 20, and the percent of male mice in Experiment 21, that walked first toward and fed first on the bait composition with dry-rendered lard and cracklings (treatment bait according to an embodiment of the invention) or Provoke (control bait). The composition of the new six- component bait was identical to that described in the caption of FIGURE 12 except that it was diluted by 1/100. Percent first-choice and percent first-feeding data were analyzed by χ test with Yates correction. In Experiment 20, bars with an asterisk (*) indicate a significant preference for the treatment bait (P≤ 0.05). DETAILED DESCRIPTION
Throughout the following description, specific details are set forth in order to provide a more thorough understanding of the invention. However, the invention may be practiced without these particulars. In other instances, well known elements have not been shown or described in detail to avoid unnecessarily obscuring the invention. Accordingly, the specification and drawings are to be regarded in an illustrative, rather than a restrictive, sense. The present inventors have taken an innovative approach for developing replacement rodent baits, namely to identify key semiochemicals in various foodstuffs attractive to mice and rats, to separate them from the potentially allergenic components in the foodstuffs, and to formulate these semiochemicals as novel baits. These synthetic semiochemical baits can be used alone or in combination with whole foodstuffs to make a more complete bait. Two such foodstuffs that pertain to this invention are dry-rendered lard and cracklings. Lard is the melted and clarified fat of pigs. There are three common kinds of lard: leaf lard made from the fatty deposit surrounding the kidneys of a pig, fatback lard made from the subcutaneous fat on the back of a pig, and soft caul lard made from fat surrounding the digestive organs.
However, lard can be made from any fatty tissue of a pig, e.g. cheeks cut from
slaughtered animals. Lard can be wet-rendered by boiling fatty tissue, or dry-rendered by heating fatty tissue in the absence of water. Hereafter, wet-rendered lard is referred to as lard, unless clarification is needed to ensure that it is distinguished from dry-rendered lard. Cracklings are the crisp residue left after lard has been dry-rendered.
The following examples are non-limiting embodiments of the invention. EXAMPLE 1
Identification and selection of candidate bait components from common foodstuffs
Food sources known to be highly attractive to mice and rats were acquired, including various types of candy, fruit, cheese, and nuts. Their headspace volatiles were collected and potentially attractive components therein were identified by various analytical procedures, including (but not limited to) gas chromatography (GC), coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry. Identification of compounds was confirmed by comparing their GC retention times and mass spectra with those of authentic standards, which were purchased or synthesized. In selecting candidate semiochemicals in these very complex volatile mixtures for formulation in baits, the present inventors were guided by (/) antennal responses to compounds by cockroaches as substitute generalist omnivores, (/'/') the re-occurrence of compounds in several foodstuffs, and (in) the olfactory sensation of our own noses, assuming that the noses of humans and rodents can perceive the same compounds.
Taking this approach, the following candidate attractive semiochemicals for mice and rats were selected: maple lactone (2-hydroxy-3-methylcyclopent-2-en-l-one) from candy; strawberry furanone (4-hydroxy-2,5-dimethylfuran-3-one) from strawberries; butyric acid, 2,3-butadione, and 3-methylbutanal from Swiss cheese; and 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one and γ-octalactone from hazelnuts.
EXAMPLE 2
General design for T-tube olfactometer experiments
The present inventors tested behavioral responses of ( ) >12-week-old reproductively active adult male and female rats (Rattus norvegicus), and (/ ) >8-week-old
reproductively active adult male and female mice (Mus musculus), to semiochemical baits in a two-choice T-tube olfactometer (FIGURE 1). The experimental set-up consisted of 3 glass aquaria (30 x 30 * 60 cm each) interconnected by a T-tube 3 (75 χ 50 cm; 10 cm diam). It was housed in a beige enclosure (3 2 1.3 m; not shown) with a small observation port. For each replicate, a single rodent was deprived of food (but not water) for 8 h, and then placed into aquarium 1 which was illuminated by a 20-W red bulb to facilitate observations of the animal's position. Following 1 h of acclimation aluminum gate 2 was opened and the animal allowed to enter the stem of T-tube 3 in response to semiochemical baits randomly assigned to live trap 5a or 5b in the outer corner of aquaria 4a or 4b.
The first choice of side chamber 4a or 4b by the test animal, the time (up to 30 min) it spent in 4a or 4b, and the trap it entered were recorded. The animal was classed as a responder if it entered with all four paws into aquarium 4a or 4b. Between each replicate, aquaria were cleaned with a dish soap detergent and wiped with a commercial pet urine odour remover (Nature's Miracle®, Eight in One Pet Products, Haupauge, NY, USA). The T-Tube was cleaned with Sparkleen and baked at 200° C for 20 min. The first-choice and trapped data were analyzed using a binomial test, and time-spent data was analyzed using a Students t test. In all cases, a = 0.05. EXAMPLE 3
Attraction of mice to synthetic maple lactone
In Experiment 1, the efficacy of synthetic2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone), a sweet-smelling volatile constituent of scotch pudding, formulated in wet-rendered lard (Tenderflake; Maple Leaf Foods, Mississauga, Ontario, Canada) (treatment bait) was tested in attracting mice. Compared to lard alone (control bait), mice walked significantly more often first toward the treatment bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 2). EXAMPLE 4
Attraction of mice to Swiss cheese or synthetic maple lactone
In Experiment 2, the attractiveness of Swiss cheese added to lard (treatment bait) was tested against attractiveness of synthetic 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone) formulated in lard (control bait).
There was no significant difference in the number of mice that walked first toward the treatment or the control bait, the time they spent near the treatment or control bait, and the number of times mice entered traps with a treatment or control bait (FIGURE 3). Therefore, synthetic maple lactone and cheese are equally attractive to mice.
EXAMPLE 5
Attraction of mice to two- or three-component baits Experiments 3-5 explored potential additive or synergistic effects of several components formulated in lard on the attraction of mice.
Experiments 3 and 4 tested whether two-component baits are more effective in attracting mice than maple lactone as a single-component bait. Experiment 3 compared the response of mice to synthetic 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone) alone or with synthetic 4-hydroxy-2,5-dimethylfuran-3-one (strawberry furanone), and Experiment 4 compared the response of mice to synthetic maple lactone alone or with cheese. In both Experiments 3 and 4, mice unexpectedly walked significantly more often first toward the two-component baits, spent significantly more time near them, and entered significantly more often the traps baited with them (FIGURE 4). Considering that two-component baits in Experiments 3 and 4 proved more attractive to mice than single-component baits, Experiment 5 compared the response to a three- component bait (synthetic maple lactone plus synthetic strawberry furanone plus cheese) with that to a two-component bait (synthetic maple lactone plus synthetic strawberry furanone).
In Experiment 5, mice surprisingly walked significantly more often first toward the three- component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 4).
These results indicate that mice are significantly more attracted to complex multi- component food odors than to simple ones.
EXAMPLE 6
Attraction of mice to three-, four- or five-component baits Taking into account that a three-component bait in Experiment 5 was more attractive to mice than a two-component bait, Experiment 6 tested whether an even more complex five-component bait, comprising synthetic 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone), three synthetic cheese volatiles (CV) (butyric acid, 2,3-butadione, and 3- methylbutanal) and ground-up hazelnut, was more attractive to mice than the same bait lacking hazelnut. In Experiment 6, mice walked significantly more often first toward the five-component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 5). The cheesy but somewhat offensive smell of butyric acid prompted us to test whether butyric acid was an absolutely essential component of the bait. In Experiment 7, three- and four-component baits were identical to those in Experiment 6, except that both lacked butyric acid. In Experiment 7, mice walked significantly more often first toward the four-component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 5).
These results support the conclusion that mice are strongly attracted to complex odorants, particularly those that include volatile components of hazelnuts or other nuts that contain similar components. The deletion of butyric acid without diminishing the degree of attraction suggests that some degree of redundancy occurs among the components of complex odorants. EXAMPLE 7
Synthetic hazelnut volatiles in lieu of ground-up hazelnut
In Experiments 6 and 7, hazelnut as part of a complex bait greatly contributed to the attraction of mice, but non-volatile components are allergenic to some people and pets. Thus, Experiments 8 and 9 tested whether hazelnut could be replaced by two or three non-allergenic key hazelnut volatiles (HV) (6-methyl-4-heptanone; 6-methyl-(E)-2- hepten-4-one; γ-octalactone) without diminishing the synergistic effect of hazelnut. In both Experiments 8 and 9, mice walked more often first toward the bait containing two (Experiment 8) or three (Experiment 9) synthetic hazelnut volatiles, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 6).
This result suggests that mice are highly responsive to volatiles that indicate the presence of nutritious hazelnuts, as well as other nuts that contain similar volatile constituents.
EXAMPLE 8
Effect of six-component bait on capture of rats
To determine whether the six-component bait, which so effectively attracted and prompted capture of mice in Experiment 9, would be similarly effective in attracting and prompting capture of rats, Experiment 10 tested the response of rats {Rattus norvegicus) to the same six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en- 1 -one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone.
In Experiment 10, rats unexpectedly walked significantly more often first toward the six- component bait formulated in lard than to lard alone (FIGURE 7). They also spent significantly more time near the six-component bait, and entered significantly more often the trap baited with it (FIGURE 7).
EXAMPLE 9
Effect of semiochemical dose on attraction and capture of rodents
The dose of odorants emanating from baits affects both the space over which animals are attracted and the proportion of them entering a trap. A higher dose may be more effective in long-range attraction of animals but may become too overwhelming to mediate short-range orientation towards, and entry into, a trap. In Experiments 11 and 12, a ten-fold lower dose of the six-component bait comprised of 2-hydroxy-3- methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6- methyl-(E)-2-hepten-4-one, and γ-octalactone was tested to see if it would retain its effectiveness against mice (Experiment 11) and rats (Experiment 12).
In Experiments 1 1 and 12, mice and rats walked more often first toward the low-dose six- component bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 8). Thus the dose could be lowered by without affecting the positive response of both mice and rats.
EXAMPLE 10
Comparative attractiveness of the six-component bait against the leading commercial baits
To gauge the relative attractiveness and operational potential of the six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone (see EXAMPLE 9), Experiment 13 tested the six-component bait with mice (Mus musculus) against
Provoke® Mouse Attractant, and Experiment 14 tested the six-component bait with rats (Ratt s noeigicus) against Provoke® Professional Rat Attractant.
Compared to the response to Provoke® Mouse Attractant, mice in Experiment 13 walked more often first toward the bait, spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 9). In Experiment 14, rats walked significantly more often first toward the six-component bait, spent significantly more time near it, and entered more often the trap baited with it (FIGURE 9).
Therefore, it is unexpectedly shown that a novel complex composition comprised of synthetic volatiles from three different foodstuffs, is more attractive to mice {Mus musculus) and rats (Rattus noregicus) than are Provoke® Mouse Attractant and
Provoke® Professional Rat Attractant, respectively.
EXAMPLE 11
Comparative attractiveness of a modified six-component bait against the leading commercial bait for mice
To reduce the potential cost of an operational six-component bait, 6-methyl-4-heptanone was removed and replaced with butyric acid to form a modified (new) six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, butyric acid, 2,3-butadione, 3- methylbutanal, 6-methyl-(£)-2-hepten-4-one, and γ-octalactone.
In Experiment 15, said new six-component bait was tested with mice {Mus musculus) against Provoke® Mouse Attractant, and for comparison in Experiment 16, the original six-component bait comprised of 2-hydroxy-3-methylcyclopent-2-en-l-one, 2,3- butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ- octalactone was also tested with mice {Mus musculus) against Provoke® Mouse
Attractant.
Unexpectedly in Experiment 15 the response of mice to the new six-component bait was almost identical to the response of mice to the original six-component bait in Experiment 16 (FIGURE 10). In both cases significantly more mice were trapped in traps baited with the six-component bait than in traps baited with Provoke® Mouse Attractant. However, the new six-component bait did not induce a significantly greater first choice by mice than occurred to Provoke® Mouse Attractant (FIGURE 10). The results of Experiments 15 and 16 indicate that baits of different compositions can be used with similar effectiveness. These results reinforce support of the hypothesis that there is some redundancy among the components of complex odorants for rodents, and indicate that baits of different compositions can have comparable operational potential. EXAMPLE 12
Effect of body odorants on bait attractiveness
Although the original six-component blend was already more effective than Provoke® Mouse Attractant and Provoke® Rat Attractant in capturing mice and rats in Experiments 13 and 14 (see EXAMPLE 10), an additional experiment was conducted to determine whether the effectiveness of the six-component bait could be increased by adding body odor components (pheromones) of mice and rats. This approach took into account that traps which emanate the body scent of previously captured mice or rats are more effective in capturing mice or rats than are new or thoroughly cleaned traps.
Body swabs were taken from lactating females and infants of both house mice (Mus musculus) and Norway rats (Rattus norvegicus) and analysed by coupled gas
chromatography-mass spectrometry. These analyses revealed that the chemical composition of all body swabs was very similar comprising nine constituents, as follows: nonanoic acid (1.2%), decanoic (capric) acid (2.4%), dodecanoic (lauric) acid (17.8%), tetradecanoic (myristic) acid (5.9%), hexadecanoic (palmitric) acid (41.4%), (Z)-octadec- 9-enoic (oleic) acid (17.8%), octadecanoic (stearic) acid (5.9%), lactic acid (1.8%) and glycerol (5.9%). Experiment 17 was done to determine the effect of these body odor components on enhancing the attractiveness of the new six-component blend (see EXAMPLE 11). Two representative components, lactic acid and oleic acid, were selected and formulated at a 1 :9 ratio in wet-rendered lard at 1% in combination with the new six-component bait.
In Experiment 17, female mice walked significantly more often first toward the new six- component bait with oleic and lactic acid (pheromone constituents) added than they did to the new six-component bait alone (FIGURE 11). They also spent significantly more time near it, and entered significantly more often the trap baited with it (FIGURE 11), clearly indicating that body odorants enhance bait effectiveness.
The results of Experiment 17 further support our findings that rodents are attracted by complex chemical stimuli, and demonstrate that combining body odor pheromone with a food bait will enhance attraction to a level greater than that achieved by a food bait alone.
EXAMPLE 13
Search for feeding stimulants
When the new six-component bait plus the two-component body odour blend was tested in a field experiment with snap traps, not as many mice as anticipated were captured and killed. The present inventors hypothesized that this was because the mice did not attempt to feed on, and thus disturb, the bait to an extent needed to trigger the snapping mechanism. The present inventors therefore searched for an effective feeding stimulant that could be added to the new six-component bait, with or without body odor constituents. Many feeding trials were run in the laboratory with a wide range of carefully selected food types, including oats, honey oats, wheat, barley, rice, peas, lentils, beans, corn, starch, pudding, molasses, gelled peanut oil, baby food, pork rinds, beef tallow, dry-rendered lard, and cracklings. In each replicate of these feeding trials, a house mouse was offered four or five types of food. Each food type was placed in a Petri dish, randomly assigned to a corner or the centre of a medium-size aquarium, and weighed before and after the trial to determine the amount which was consumed by the mouse.
Unexpectedly, dry-rendered lard and cracklings were clearly the most preferred food types. Therefore, they were selected for admixture to the new six-component bait (see EXAMPLE 11). EXAMPLE 14
Test of adding dry-rendered lard and cracklings to the new six-component bait on feeding by mice and rats
To determine whether adding dry-rendered lard and cracklings to the new six-component bait indeed stimulated feeding by mice and rats a composition comprising the new six- component bait (50%), dry-rendered lard (40%) and cracklings (10%) was prepared. The present inventors then tested whether house mice (Experiment 18) and rats (Experiment 19) preferred to feed on this composition or on a composition containing only the new six-component bait.
In Experiment 18, female house mice unexpectedly consumed significantly more bait containing dry-rendered lard and cracklings than the new six-component bait without these constituents (FIGURE 12). Similarly, in Experiment 19, female rats also consumed significantly more bait containing dry-rendered lard and cracklings than the bait without these ingredients (FIGURE 12).
These results confirm the feeding stimulatory effect of dry-rendered lard and cracklings on mice (see EXAMPLE 13), demonstrate the same effect on rats, and show that greater feeding stimulation can be achieved by a composition that includes synthetic food constituents, dry-rendered lard and cracklings than can be achieved by the synthetic food composition alone. EXAMPLE 15
Attraction and feeding response of house mice to a very low dose of the new six- component blend in combination with dry-rendered lard and cracklings
In Experiment 15 (Figure 10) the new six-component bait did not induce greater attraction (first choice) in the T-tube olfactometer than was achieved by Provoke®
Mouse Attractant. Experiments 18 and 19 (Figure 12) showed that adding dry-rendered lard and cracklings to the new six-component bait enhanced feeding over that induced by the new six-component bait alone. This final example tested whether adding dry- rendered lard and cracklings would also enhance the attractive properties of the new six- component bait.
Experiment 20 tested a reduced dose (1/100 dilution) of the new six-component bait in combination with dry-rendered lard and cracklings, with Provoke® Mouse Attractant as a control stimulus. The composition of the experimental bait was: the new six-component blend (1/100 dilution) (50%), dry-rendered lard (40%) and cracklings (10%). The experiment was run in the T-tube olfactometer (see EXAMPLE 2, FIGURE 1).
In Experiment 20, female house mice surprisingly walked significantly more often first toward the composition bait than toward Provoke® Mouse Attractant, and also fed first significantly more often on the composition bait (FIGURE 13). Similarly, in Experiment 21, male house mice walked more often first toward the composition bait than toward Provoke, and fed more often on the composition bait than on Provoke® Mouse
Attractant. The results of Experiments 18-21 unexpectedly show that combining a synthetic food bait (the new six-component bait) with naturally derived foodstuffs (dry-rendered lard and cracklings) enhance both attraction and feeding stimulation of rodents. As will be apparent to those skilled in the art in the light of the foregoing disclosure, many alterations and modifications are possible in the practice of this invention without departing from the spirit or scope thereof.
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Claims

WHAT IS CLAIMED IS:
1. A composition used in effective amount to attract and induce feeding by mice and rats comprising at least one compound derived from a source selected from the group consisting of sugar-containing confections, strawberries, cheese, nuts, lactating female rodents and infant rodents.
2. The composition of claim 1, wherein the sugar-containing confections is selected from the group consisting of butterscotch candy, pudding, and cookies.
3. The composition of claims 1 or 2, wherein the compound derived from sugar- containing confections comprises 2-hydroxy-3-methylcyclopent-2-en-l-one (maple lactone).
4. The composition of any one of claims 1 to 3, wherein the compound derived from strawberries comprises 4-hydroxy-2,5-dimethylfuran-3-one (strawberry furanone).
5. The composition of any one of claims 1 to 4, wherein the cheese comprises Swiss cheese.
6. The composition of any one of claims 1 to 5, wherein the compound derived from the cheese is selected from the group consisting of butyric acid, 2,3-butadione, and 3-methylbutanal.
7. The composition of any one of claims 1 to 6, wherein the nuts can comprise hazelnuts.
8. The composition of claim 7, wherein the compound from hazelnuts is selected from the group consisting of 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, and γ-octalactone.
9. The composition of any one of claims 1 to 8, wherein the lactating female rodents or infant rodents are mice or rats.
10. The composition of any one of claims 1 to 9, wherein the compound from the lactating female rodents or infant rodents females is selected from the group consisting of nonanoic acid, decanoic (capric) acid, dodecanoic (lauric) acid, tetradecanoic (myristic) acid, hexadecanoic (palmitic) acid, (Z)-octadec-9-enoic (oleic) acid, octadecanoic (stearic) acid, lactic acid and glycerol.
1 1. A synthetic composition used in effective amount to attract and induce feeding by mice and rats, wherein said composition is comprised of two or more compounds selected from the group consisting of 2-hydroxy-3-methylcyclopent-2-en-l-one, 4-hydroxy-2,5-dimethylfuran-3-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, γ-octalactone, nonanoic acid, decanoic (capric) acid, dodecanoic (lauric) acid, tetradecanoic (myristic) acid, hexadecanoic (palmitic) acid, (2)-octadec-9-enoic (oleic) acid, octadecanoic (stearic) acid, lactic acid and glycerol.
12. A synthetic composition according to claim 11 comprising 2-hydroxy-3- methylcyclopent-2-en-l-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6- methyl-(E)-2-hepten-4-one, γ-octalactone, (Z)-octadec-9-enoic (oleic) acid and lactic acid.
13. The composition of claim 12, wherein the proportions by weight of the
components are: 2-hydroxy-3-methylcyclopent-2-en-l-one (0.001-50%), butyric acid (0.001-50%), 2,3-butadione (0.001-50%), 3-methylbutanaI (0.001-50%), 6- methyl-(E)-2-hepten-4-one (0.001-50%), γ-octalactone (0.001-50%), (Z)-octadec- 9-enoic (oleic) acid (0.001-50%) and lactic acid (0.001-50%).
14. A composition used in effective amount to attract and induce feeding by mice and rats, wherein said composition is comprised of two or more compounds selected from the group consisting of 2-hydroxy-3-methylcyclopent-2-en-l-one, 4- hydroxy-2,5-dimethylfuran-3-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, γ-octalactone, nonanoic acid, decanoic (capric) acid, dodecanoic (lauric) acid, tetradecanoic (myristic) acid, hexadecanoic (palmitic) acid, (Z)-octadec-9-enoic (oleic) acid, octadecanoic (stearic) acid, lactic acid and glycerol, combined with one or more of dry- rendered lard and cracklings produced during a dry rendering process.
15. A composition according to claim 14 comprising 2-hydroxy-3 -methyl cyclopent- 2-en-l-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6-methyl-(E)-2-hepten- 4-one, γ-octalactone, and dry-rendered lard and cracklings produced during the dry rendering process.
16. The composition of claim 15, wherein the proportions by weight of the
components are: 2-hydroxy-3-methylcyclopent-2-en-l-one (0.001-50%), butyric acid (0.001-50%), 2,3-butadione (0.001-50%), 3-methylbutanal (0.001-50%), 6- methyl-(E)-2-hepten-4-one (0.001-50%), γ-octalactone (0.001-50%), and dry- rendered lard (0.001-50%) and cracklings produced during the dry rendering process (0.001-50%).
17. A method of attracting and inducing feeding by mice and rats by using in
effective amount a composition comprised of compounds derived from a source selected from the group consisting of sugar-containing confections, strawberries, cheese, nuts, lactating female rodents and infant rodents.
18. The method of claim 17, wherein the compounds are synthetic.
19. The method of claim 17 or 18, wherein the composition comprises two or more compounds selected from the group consisting of 2-hydroxy-3-methylcyclopent- 2-en-l-one, 4-hydroxy-2,5-dimethylfuran-3-one, butyric acid, 2,3-butadione, 3- methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, γ- octalactone, nonanoic acid, decanoic (capric) acid, dodecanoic (lauric) acid, tetradecanoic (myristic) acid, hexadecanoic (palmitic) acid, (Z)-octadec-9-enoic (oleic) acid, octadecanoic (stearic) acid, lactic acid and glycerol.
20. The method of claim 17, wherein the synthetic composition used in effective amount to attract and induce feeding by mice and rats comprises 2-hydroxy-3- methylcyclopent-2-en-l-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6- methyl-(E)-2-hepten-4-one, γ-octalactone, (Z)-octadec-9-enoic (oleic) acid and lactic acid.
21. The method of claim 17, wherein the synthetic composition used in effective amount to attract and induce feeding by mice and rats comprises two or more compounds selected from the group consisting of 2-hydroxy-3-methylcyclopent- 2-en-l-one, 4-hydroxy-2,5-dimethylfuran-3-one, butyric acid, 2,3-butadione, 3- methylbutanal, 6-methyl-4-heptanone, 6-methyl-(E)-2-hepten-4-one, γ- octalactone, nonanoic acid, decanoic (capric) acid, dodecanoic (lauric) acid, tetradecanoic (myristic) acid, hexadecanoic (palmitic) acid, (Z)-octadec-9-enoic (oleic) acid, octadecanoic (stearic) acid, lactic acid and glycerol, combined with one or more foodstuffs selected from the group consisting of dry-rendered lard and cracklings produced during the dry rendering process. The method of claim 17, wherein the synthetic composition used in effective amount to attract and induce feeding by mice and rats comprises 2-hydroxy-3- methylcyclopent-2-en-l-one, butyric acid, 2,3-butadione, 3-methylbutanal, 6- methyl-(E)-2-hepten-4-one, γ-octalactone, dry-rendered lard and cracklings produced during the dry rendering process.
The method of any one of claims 17 to 22, wherein the synthetic composition and foodstuffs used in effective amount to attract and induce feeding by mice and rats is formulated into a paste, gel, powder or pellet.
The method of any one of claims 17 to 22, wherein the synthetic composition and foodstuffs used in effective amount to attract mice and rats is used in a trap.
The method of any one of claims 17 to 22, wherein the synthetic composition and foodstuffs used in effective amount to attract mice and rats is formulated into a bait laced with a rodenticide.
The method of any one of claims 17 to 25, wherein the mice can be Mus musculus.
The method of any one of claims 17 to 25, wherein the rats can be Rattus norvegicus.
PCT/CA2012/000638 2011-07-04 2012-06-28 Compositions and methods for attracting and stimulating feeding by mice and rats WO2013003946A1 (en)

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