Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
  • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
  • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
  • A61K31/201—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having one or two double bonds, e.g. oleic, linoleic acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/20—Hypnotics; Sedatives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/22—Anxiolytics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/26—Psychostimulants, e.g. nicotine, cocaine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/10—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4
  • A61P5/16—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4 for decreasing, blocking or antagonising the activity of the thyroid hormones
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid

Definitions

• the present invention relates to a composition
• a composition comprising a mixture of fatty acids or derivatives thereof derived from secretions of the uropygial glands of birds.
• This composition named an avian appeasing pheromone, can be used to decrease stress, anxiety and aggressiveness in birds.
• Stress is associated with an objective aggression and the consequences appear in altering different systems such as humoral, metabolic, immunolgic and/or behavioral systems.
• the hormonal response is traditionally determined by a release of cortisol, a decrease in the secretion of growth hormone and an increase in thyroid hormones and also the sexual steroids. Stress also has a role for diminishing either directly or indirectly the amount of consumption, which is a principal problem with animal breeders. This decrease in the amount of consumption in animals can be caused either by an increase in metabolism or a decrease in food consumption due to stress.
• Physiological stressors can also result in an abnormally elevated corticosterone secretion. Campo J.L., S.G. Davila,. Poult. Sci. 81 : 1637-1639. (2002); Moberg G.P.,. In : The Biology of animal stress (Moberg & Mench Ed.), CABI, 1-23(2000) Besides physiological tests, behavioral tests can also be performed to evaluate stress and/or fear in avians. One of the most utilized behavioral tests is the test of Tonic immobility (Tl) as described by Campo J.L., S.G. Davila,. Poult. Sci. 81 : 1637-1639 (2002).
• Tl Tonic immobility
• Litter In a confinement housing system, the birds remain indoors and are provided with light twenty-four hours a day to encourage feed consumption. Litter is generally spread on the floor in the hen houses which acts to absorb moisture, dilute manure and provides cushioning and insulation for the birds. In the conventional industry, litter is spread 2 to 4 inches deep and maintained at a 20% to 30% moisture content. There is generally a rest period of about a week between flocks, when the hen houses are cleaned out of de-caked. By definition, pheromones are substances released by the body that cause a predictable reaction by another individual of the same species.
• a number of different glands are known to produce pheromones in male mammals such as the submaxillary salivary glands, the parathyroid glands and the sebaceous glands. Pheromones that are secreted in submaxillary salivary and parathyroid glands in males, are used to mark females during courtship. In boars, the secretion of these glands results in agnostic behavior. These secretions are known to contain a mixture of androstenol and andosterone.
• pheromones in mammals can be used to reduce stress, anxiety and aggressiveness as demonstrated in U.S. Patent Nos. 6,077,87, 6,054,481 and 6,169,113. These pheromones were derived from secretions of mammalian mammary glands. However, mammary glands do not exist in birds. Moreover, there are some doubts among specialists about the presence of pheromones in birds since there is no vomeronasal organ, which is a sensory organ with its own pathway to the brain.
• the uropygial glands in birds are a bilobed holocrine gland with secretions that form several functions in birds. These functions include waterproofing the feathers, manufacturing of vitamin D precursors, keeping the bill, skin and feathers supple and preventing bacterial infection.
• the secretions of the uropygial gland are generally spread over the feathers during the act of preening and thus weatherproofs the feathers.
• Bohnet et al in the Journal of Biochemistry vol. 226, No. 15 pp. 9795- 9804 (1991 ) describe the production in female mallards of sex pheromones, diesters of 3-hydroxy fatty acids in their uropygial glands during the mating season. Outside the mating season the uropygial gland only produced a usual monoester wax. It is not known how the uropygial gland switches its biological processes to produce sexual pheromones during the mating season only.
• an aspect of the present invention is to provide an avian appeasing pheromone derived from the uropygial gland of an avian.
• the present invention provides a composition
• a composition comprising an avian appeasing pheromone comprising lauric acid, palmitic acid, linoleic acid and oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the present invention provides a composition
• an avian appeasing pheromone comprising a mixture of about 12.3 to 13.7 (w%/w%) of lauric acid, about 38.0 to 42.0 (w%/w%) palmitic acid, about 32.3 to 35.7 (w%/w%) linoleic acid and about 12.0 to 14.0 (w%/w%) oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the present invention provides a process to treat stress in an avian, said process comprising administering to an avian in need of such treatment an avian appeasing pheromone derived from secretions around the uropygial gland of an avian.
• the present invention relates to a process of treating weight loss in an avian, said process comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• the present invention provides a process of treating a domestic avian during transportation to eliminate their anxiety, said process comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• the present invention provides a process to improve feed conversion in an avian comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• the present invention provides a composition comprising an avian appeasing pheromone derived from around the uropygial gland of an avian.
• the present invention discloses a composition
• a composition comprising an avian appeasing pheromone comprising lauric acid, palmitic acid, linoleic acid and oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the present invention provides a composition
• a composition comprises an avian appeasing pheromone comprising about 12.3 to 13.7 (w%/w%) of lauric acid, about 38.0 to 42.0 (w%/w%) palmitic acid, about 32.3 to 35.7 (w%/w%) linoleic acid and about 12.0 to 14.0 (w%/w%) oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the derivatives of the present invention include esters such as methyl esters, salts, alcohols, ketones, ethers, aldehydes, sterols and amides of lauric acid, palmitic acid, linoleic acid and oleic acid.
• the present invention provides a process for the treatment of stress in an avian said process comprising administering to an avian in need of such treatment a composition comprising an avian appeasing pheromone derived from secretions around the uropygial gland of an avian.
• the present invention discloses a process of treating weight loss in an avian, said process comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• the present invention provide a process of treating a domestic avian during transportation to eliminate their anxiety, said process comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• the present invention discloses a process to improve feed conversion in an avian comprising administering to an avian in need of such treatment an avian appeasing pheromonal composition derived from secretions around the uropygial gland of an avian.
• Fig. 1 are gas chromatograph/mass spectroscopy spectrum profiles of hens that were not followed by their chicks and hens that were followed by the chicks at day 8.
• Fig. 2 are gas chromatograph/mass spectroscopy spectrum profiles of hens that were followed by their chicks at days 4, 8 and 12.
• Fig. 3 is a bar graph showing that gain in weight of young chicks. ⁇ indicates that the chicks were treated with avian appeasing pheromone, while ⁇ indicates the chickens were treated with a placebo.
• Fig. 4 is a graph showing the increase in weight gain in young chicks when administered avian appeasing pheromone (top line) in comparison with a placebo (bottom line).
• Fig. 5 is a bar graph showing the levels of cortisol in hens (Females) and roosters (Males) and males + females combined (M+F).
• Fig. 6 is a bar graph showing the T4 levels in hens (Females) and roosters (Males) and males + females combined (M+F).
• Fig. 7 is a bar graph showing the cortisol levels in chickens under different housing conditions . ⁇ indicates that the chickens were treated with avian appeasing pheromone, while ⁇ indicates the chickens were treated with a placebo.
• Fig. 8 is a bar graph showing the T4 levels in chickens under different housing conditions . ⁇ indicates that the chickens were treated with avian appeasing pheromone, while ⁇ indicates the chickens were treated with a placebo.
• Fig. 9 is a bar graph showing the weight gain in chickens under different housing conditions . ⁇ t indicates that the chickens were treated with avian appeasing pheromone, while ⁇ indicates the chickens were treated with a placebo.
• Fig. 10 is a bar graph showing the total cortisol levels under different housing conditions.
• Fig. 1 1 is a bar graph showing the ratio of H/L on hens on different dates of the same year 2002.
• the word “bird” and “avian” are used interchangeably and encompass any warm-blooded animal with feathers and wings, that lays eggs and is usually able to fly. Examples of birds, include, but are not limited to chicks, chickens, guinea hens, parrots, turkeys, hens, ducks, geese and the like.
• stress is meant the reaction of an animal body to forces of deleterious nature, infection and various abnormal states that tend to disturb homeostasis. This reaction may be a physical reaction or an emotional reaction including anxiety.
• anxiety is meant an apprehension of anger and dread accompanied by restlessness, tension and the like, which is a reactional status characterized by a high probability to provide behavioral and emotional responses of fright. In neurophysical terms, this anxious state is accompanied by an hyperactivity of the noradrenergic and serotonin systems.
• pheromone is meant a substance released by the body of a particular species that causes a predictable reaction by another individual of the same species, which substance may serve, for example, as a specific attractant, social communicator, sexual stimulant and the like.
• pheromone By “appeasing pheromone” is meant a pheromone that calms, soothes or relieves stress, anxiety and aggressiveness. Appeasing pheromones should be distinguished from sexual pheromones, the latter that act as either sexual stimulants or attractants.
• stress-associated diseases any disease whose symptoms increase due to stress.
• improved feed conversion means the reduction of the ratio in food consumption/ weight gained.
• solution is meant a solid that is dispersed through a liquid either by being dissolved in it or being in suspension.
• an active pheromonal composition that is species-species specific in birds and which can be used to enhance or act synergistically with the basic pheromonal composition to increase the effectiveness in specific species of the composition.
• heterophile all white blood cells that are not lymphocytes that are found in avians.
• isomers includes structural isomerism and spatial isomerism and refers to the isomers of the fatty acids of lauric acid, palmitic acid, linoleic acid and oleic acid, as well as their derivatives.
• the composition can include, for example only, lauric acid, a derivative of palmitic acid, an isomer of linoleic acid and an isomer of a derivative of oleic acid. It will be appreciated that the compositions of the present invention encompass all of the permutations (and not only that exemplified above) of the mixtures of the disclosed fatty acids with their derivatives and/or isomers thereof. Furthermore, the derivatives and isomers referred to herein have the same weight percentages as their fatty acid counterparts. For example a derivative, isomer or derivative of an isomer for lauric acid should have a concentration between 12.3 to 13.7 (w%/w%).
• the present invention relates to the identification of an avian appeasing pheromonal composition that is derived from secretions around the uropygial gland of birds.
• compositions of the present invention are appeasing and pheromonal in origin and are made up of volatile molecules, the essential components of these molecules being amines and fatty acids from indolic derivatives, as well as esters of these amines and fatty acids.
• composition of the present invention comprises a mixture of lauric acid, palmitic acid, linoleic acid and oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the basic composition of the present invention comprises a mixture of about 12.3 to 13.7 (w%/w%) of lauric acid, about 38.0 to 42.0 (w%/w%) palmitic acid, about 32.3 to 35.7 (w%/w%) linoleic acid and about 12.0 to 14.0 (w%/w%) oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• composition can also be attached to a chemical carrier provided that the bioactive structure of the fatty acids is preserved.
• carrier molecules include, but are not limited to, resins, liposomes, crown compounds, carrier proteins, polymers and the like.
• the fatty acids can be used in their pure form, i.e., as a free fatty acid, as well as their derivative form such as esters of fatty acids or salts of fatty acids, as well as alcohols of fatty acids, ketones of fatty acids, ethers of fatty acids and amides of fatty acids.
• These fatty acid derivatives can replace one or more or all of the fatty acids in the compositions of the present invention and have the same effects.
• the derivatives of the fatty acids can be found in the compositions or in the solutions of the present invention.
• the composition has been found to have an appeasing effect in avians and can be used to relieve stress, anxiety, reduce aggressive behavior, enhance weight gain and enhance egg production.
• An enhancer composition containing between 5% to 35% (w%/w%) can also be added to the avian appeasing pheromone composition, if desired.
• This enhancer composition comprises volatile organic compounds and mixtures thereof.
• This enhance composition may be species-specific in nature and may vary according to the avian species selected for use in the present invention.
• the compounds that can be used in the enhancer composition include, but are not limited to, amines and fatty acids from indolic derivatives, esters of these amines and fatty acids, ketones such as acetone, alcohols, sterols and the like.
• Nontoxic fillers can also be added to the composition, which include fatty acids, alcohols, amines, squalene and glycerol.
• the present invention comprises the avian appeasing pheromonal composition in solution.
• the present invention thus provides a solution of lauric acid, palmitic acid, linoleic acid and oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the present invention provides in solution about 12.3 to 13.7 (w%/w%) of lauric acid, about 38.0 to 42.0 (w%/w%) palmitic acid, about 32.3 to 35.7 (w% w%) linoleic acid and about 12.0 to 14.0 (w%/w%) oleic acid and/or derivatives thereof and/or isomers thereof, as well as mixtures of one or more of these fatty acids with one or more of their derivatives and/or one or more of their isomers.
• the composition may be in the form of a solution, aerosol spray, gel, slow release matrix, shampoo and the like.
• the composition can also be placed in liposomes, in a diffuser, in polymers, is in an electric diffuser and be microencapsulated.
• the concentration of the above-mentioned fatty acids may vary depending upon the final form of use. However, the concentrations of the specific fatty acids that are utilized and their concentration may be ascertained and tested according to the methods set forth in the present invention.
• the fatty acids which are generally solid in nature, can be diluted in any nonaqueous solvent to form the solution of the present invention. Solvents such as propylene glycol, alcohol, ether, chloroform, ethanol, benzene, carbon disulfide, polysorbate, propyl alcohol, isopropanol, 2-propanol, fixed and volatile oils and the like. Combinations of these solvents can also be used.
• a combination of propylene glycol and absolute ethanol can be used as a solvent in the present invention.
• 90% to 98% propylene glycol and 2% to 10 % absolute ethanol is set forth as an example.
• 5% to 40% isopropanol and 60% to 95% propylene glycol can also be used.
• Fatty acids are commercially available from various chemical companies in solid form. However, since it is difficult to solubilize fatty acids, the fatty acid is generally added to the solvent under constant agitation and at a temperature of between 37°C to about 38°C. 37.5°C can also be used.
• compositions of the present invention can be tested for their efficacy to prevent stress in an avian.
• Well documented stressors are, for example, the weaning of birds, the transportation of birds, and the like.
• Application of the present compositions of the invention in the form of a spray, aerosol and the like in the area surrounding the stressful events results in diminution of stress as indicated by a variety of factors such as weight gain, social behavior with respect to other birds, wounds on the body, salivary cortisol and the like.
• compositions of the present invention can be diluted and applied to various objects that the birds come in contact with such as walls, cages, in the air and toys.
• present compositions in the form of solutions can also be applied directly on the birds.
• compositions were discovered after detailed analysis of the chemical composition of secretions obtained from the uropygial gland in birds that had their young following them.
• this procedure involved swabbing the are around the uropygial glands of birds that had their young following them with a sterile compress and analyzing the chemical composition of the secretions via gas chromatography/mass spectroscopy.
• This procedure is described in more detail in the examples below. In order to fully illustrate the present invention and advantages thereof, the following specific examples are given, it being understood that the same are intended only as illustrative and in nowise limited.
• the hens were rubbed around the area of the uropygial gland or tail gland with sterilized compresses several times. The compress was immediately placed in a flask containing 10 ml of dichloromethane.
• the compress was then deabsorbed in dichloromethane by agitating the flask several times. Five ml of the sample was taken out and subjected to evaporation down to 1 ml. using air.
• the detection was effectuated on impact using (EI+) at an energy of 70 eV at
• Feeding ad libitum for food and water with the food commonly used by all breeders to feed young chicks.
• Lot A received a dosage of 2% of the Avian Appeasing Pheromone comprising 13% lauric acid (w%/w%), 40% palmitic acid (w%/w%), 34% linoleic acid (w%/w%) and 13% oleic acid (w%/w%) which was placed in an electronic diffuser and placed above the young chicks in their housing.
• Lot B received a treatment with a placebo.
• the young chicks Prior to administering the avian appeasing pheromone or the placebo the young chicks were also grouped according to their weight; i.e., at day 0.
• This example measured the influence of avian appeasing pheromone in hens and roosters subjected to hard and chronic stress.
• the stressful conditions in which the hens and chickens were subjected to include bright lights, metallic noises, and electric shock.
• Two types of stressors were provoked each day for a period of 15 minutes in a random manner, which were identical for the two lots.
• Lot A was treated with the avian appeasing pheromone (AAP) comprising 13% lauric acid (w%/w%), 40% palmitic acid (w%/w%), 34% linoleic acid (w%/w%) and 13% oleic acid (w%/w%).
• Lot B was given a placebo.
• factors were then tested such as whether the hens or roosters gained weight, the changes in cortisol levels and the changes in T4 levels using standard tests for cortisol and T4, 8 days after the end of the experiment. A difference in weight was not significantly observed. This could be because the stressors were too violent and that the pheromone could not prevent this type of violent stress.
• Lot 1 (administered avian appeasing pheromone) vs. Lot B1 (placebo) Lot 2: A2 (administered avian appeasing pheromone) vs. Lot B2
• the avian appeasing pheromone that was administered comprised 13% lauric acid (w%/w%), 40% palmitic acid (w%/w%), 34% linoleic acid (w%/w%) and 13% oleic acid (w%/w%).
• a solution of the avian appeasing pheromone titrated at 2% was the dosage used in this Example. This dosage was diffused with an electronic diffuser at a rate of 1/50m 2 .
• Weight gain, cortisol levels, weight gain and indication of consumption levels and T4 levels were measured using standard tests for cortisol and T4 levels about 40 days after the end of the experiment.
• the cortisol levels, T4 levels and weight were tested about 40 days after the initial testing.
• Example 5 This protocol tests the influence of the avian appeasing pheromone on laying hens.
• the breeding techniques for laying hens are often stressful and cause a loss of eggs that are laid and also the squabbles between individual hens.
• Starting flock 4 months after birth (year 1 )
• Starting feather pecking 8 months after birth (year 2)
• Treatment for mites Dermatysuss or Ornithonyssus:9 months after birth (year 2)
• Zoological technology was noted throughout the calendar of events described above using the following technical criteria: the rate of hens outside of their nests; the rate of hens in mass; the rate of mortality; and the alimentary consumption for those hens which were administered avian appeasing pheromone and those that were not administered avian appeasing pheromone (control).
• Mites were also tested for during the course of this example. At day 0, the first diffusers of the avian appeasing pheromone were posed and the sampling of mites was effectuated. For the 12 samples that were taken, 5 samples permitted the identification of the species of mites in the breeding; 3 were utilized and placed in a mite culture and recuperated at the end of the internal experiment; and 4 served to test the sensitivity of parasites to the mites used.
• the parasites were thus exposed to a variety of concentrations of miticide.
• the dosage was determined by the value of the notice of dosage recommendation on the product. At this dose one does not obtain the LD 50 (50% of the mites exposed over a period of 24 hours are killed, nor the LD 100 (100% of mites are killed in a 24 hour period). The LD 50 was not obtained with a dose that was 8 times superior than that recommended. The LD 100 was not obtained with a does that was 32 times superior than that prescribed.
• T1 and T2 Under experimental conditions 2 lots (T1 and T2) were subjected to the tests.
• the T1 lot corresponded to a universal test, which is not authorized in breeding since Ivermectine was used.
• Lot 2 corresponded to a test made with Amitraze.
• the LD 100 was obtained with the doses prescribed on the notices of the products, in surroundings corresponding to the conditions of breeding. The results are shown in Table 3 below:
• T1 Amitraze (T2) 1 pd 1 pd pd: prescribed dose
• the normal physiological average for a hematocrit is 55%.
• there is a diminution in the hematorcrits which signifies there is a decrease in red blood cells in the animal. This decrease indicates that the animals were injured in some manner. This blood loss is probably due to feather pecking in their tail area, which causes bad scaring and may lead to spots of blood on the eggs.
• the white blood cell analysis showed a strong presence of granulocytes, which correlates with an infestation of parasites.
• this study shows a mean value Of 2.49 for the first set of blood samples drawn and a fall of 60% for the second and third blood samples that were drawn.
• the statistical analysis indicated in the above Table 6 shows a difference between the first samples drawn and the second samples drawn and the first samples drawn and the third samples drawn but not a statistical difference between the second and third samples that were drawn.
• the mean average for breeders is 0.42 ⁇ 0.13 (3).
• the avian appeasing pheromone decreases stress in avians. However, certain conditions must be maintained such that it works effectively. The avian appeasing pheromone should be used on young chicks immediately after they hatch.
• P010 an avian appeasing pheromone comprising a mixture of 13% lauric acid methylester (w%/w%), 40 % palmitic acid methylester (w%/w%), 34.0 % linoleic acid methylester (w%/w%) and 13.0 w% oleic acid methyl ester (w%/w%) was fabricated as a slow releasing block by methods known in the art that was protected by a plastic envelope pierced with holes. The treatment consisted of a passive diffusion of P010 into the local atmosphere.
• Each block weighed 150 g and 18 blocks containing a concentration of 2% of the above avian appeasing pheromone were placed in the treated building (either P010 or a placebo). Treatment started on the day previous to the arrival of the chickens (noted as DO or Day 0). On Day 29,(D29) the blocks containing P010 were replaced with new ones. In this trial, building 01 was treated with the placebo and building 02 with P010. The treated building will be referred to hereafter as "P010" and the other one "control" (placebo). The trial was run double blinded.
• Physiological tests were performed on blood samples taken from the chickens on the day of departure to the slaughterhouse. A total of 200 of these tests were performed (50 males and 50 females were tested for each building).
• H/L Hematocrit/lymphocyte
• glucose level was measured.
• H/L was calculated using Mallacez cells on which 2 x 500 ⁇ l were spread over 50 squares (1 cm 3 ). The calculation was computed using the cross up technique. Total lymphocyte population was then counted and the ratio was computed. The hematocrit was measured after centrifugation at 9000 m/s "2 (3,500 rpm during 2 minutes) and expressed as the ratio of (total blood)/(cellular phase). The glucose level was measured using a Glucotrend®.
• BT Behavioral Test
• Each chicken was captured and placed on its back in a small hammock for a maximum of 60 seconds. Once the chicken was on its back, it is restrained gently by placing one hand touching its breast for 10 seconds and then released, while being stared at by another person eyes positioned at 1 meter, eye to eye.
• Live Weight LW
• Dead Weight DW
• Feed to Gain Ratio FGR
• Performance Index PI
• Live Weight was obtained by adding the overall weight of the chickens divided by the total number of chickens that were delivered by the breeder. The measurements were recorded separately for the male chickens and the female chickens. Dead Weight (DW) was the measurement taken of the weight of each chicken individually. The weight was then classified in 100 g classes.
• Feed to Gain Ratio was the measurement taken for the total amount of food consumed by the chickens in each building for both male and female chickens.
• Performance Index is the major index the chicken breeders took into account to evaluate performance. PI was calculated as follows: (Daily Weight Gain)/ Feed to Gain Ratio (FGR).
• the Table Yield procedure evaluates animal yield; i.e., Fat, Fillet, Leg (meat), ratio Skin/Live Weight and Global Yield: (Skin+Leg)/Live Weight.
• Total downgraded P010 male chickens (see, Table 8, above) represented 0.78% of the Dead Weight (DW), compared to 0.57% for the total downgraded control chickens (p ⁇ 0.01 ). This is due to the higher level of marked legs for the P010 treated chickens (47% versus 12%, p ⁇ 0.001 ). However, percentage of both suffocated and scratched chickens are lower in the P010 treated chickens as evidenced by the values of 1.36% (male treated) versus 1.11% male controls for suffocation and 8% (male treated) versus 22% (male control) (p ⁇ 0.001 ) for scratched chickens.
• Results concerning the female chickens are comparable to those of the male chickens (see, Table 9, above).
• the results showed a significantly higher number of marked legs for female chickens treated with P010 (8% (treated) versus 4% (control) p ⁇ 0.001), but a lower number of P010 treated female chickens suffocated (3.25% (treated) versus 6.36% (control) , p ⁇ .001 ) and were scratched (10% (treated) versus 19% (control) , p ⁇ 0.001 ) for the P010 group.
• the two observed groups showed differences among sexes for the physiological results (see Table 4 and Table 5, above). Indeed, the H/L ratio was significantly lower for the P010 chickens (p ⁇ 0.01 ) and is the unique parameter that showed a difference for female chickens among the groups. Concerning the male chickens, the Corticosterone level was higher for the control (p ⁇ 0.05), but the glucose level was lower (p ⁇ 0.01 ) for this same control group. It is interesting to observe that these results tend to show that male chickens have a level of stress that is different among the groups. The case is less for the female chickens, except for the H/L ratio, which tends to represent a social stress (Campo J.L., Davila S.G., Poult. Sci.
• BT Behavioral Test
• the P010 product reducers stress in chickens and it economically beneficial to the aviculturist as evidenced by the above results.

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