Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
  • A23L17/70—Comminuted, e.g. emulsified, fish products; Processed products therefrom such as pastes, reformed or compressed products
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
  • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
  • A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
  • A23L13/43—Addition of vegetable fats or oils; Addition of non-meat animal fats or oils; Addition of fatty acids
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
  • A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
  • A23L13/65—Sausages
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
  • A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
  • A23L13/67—Reformed meat products other than sausages
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/201—Compounds of unspecified constitution characterised by the chemical reaction for their preparation
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
  • A23L27/215—Synthetic spices, flavouring agents or condiments containing amino acids heated in the presence of reducing sugars, e.g. Maillard's non-enzymatic browning
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
  • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

• Embodiments of the invention related to the enhancement of desirable characteristics in meat products through the incorporation of beneficial fatty acids such as stearidonic acid (SDA).
• SDA stearidonic acid
• Various embodiments relate to methods of production and processing of meat products and associated food products.
• Embodiments of the present invention are directed to a method for improving foodstuffs through the utilization of plant-derived stearidonic acid ("SDA"). Specifically, the inventor provides techniques and methods for the utilization of plant-derived SDA in meat products that improve nutritional quality and palatability.
• SDA plant-derived stearidonic acid
• the fatty acids are carboxylic acids, which have from 2 to 26 carbons in their "backbone,” with none, or various numbers of unsaturated sites in their carbohydrate structure. They generally have dissociation constants (pKa) of about 4.5 indicating that in normal body conditions (physiological pH of 7.4) the vast majority will be in a dissociated form.
• Omega-3 fatty acids are long-chain polyunsaturated fatty acids (18-22 carbon atoms in chain length) with the first of the double bonds (“unsaturations") beginning with the third carbon atom. They are called “polyunsaturated” because their molecules have two or more double bonds “unsaturations” in their carbohydrate chain. They are termed "long-chain” fatty acids since their carbon backbone has at least 18 carbon atoms.
• omega-3 family of fatty acids includes alpha-linolenic acid (“ALA”), eicosapentaenoic acid (“EPA”), and docosahexaenoic acid (“DHA”).
• ALA alpha-linolenic acid
• EPA eicosapentaenoic acid
• DHA docosahexaenoic acid
• ALA is the “base” omega-3 fatty acid, from which EPA and DHA are made in the body through a series of enzymatic reactions, including the production of SDA.
• Most nutritionists point to DHA and EPA as the most physiologically important of the Omega-3 fatty acids.
• This synthesis processes from ALA are called “elongation” (the molecule becomes longer by incorporating new carbon atoms) and “desaturation” (new double bonds are created), respectively.
• ALA is primarily found in certain plant seeds (e.g., flax) while EPA and DHA mostly occur in the tissues of cold-water predatory fish (e.g., tuna, trout, sardines and salmon), and in some marine algae or microbes that they feed upon.
• cold-water predatory fish e.g., tuna, trout, sardines and salmon
• SDA Omega-3 fatty acid
• ALA ⁇ -linoleic acid
• the plant species that are specifically included within the group of those that could supply demand are: soybeans, corn, and canola, but also may include other plants as needed.
• the SDA of the invention can be used to improve the health characteristics of a great variety of food products.
• This production can also be scaled-up as needed to both reduce the need to harvest wild fish stocks and to provide essential fatty acid components for aquaculture operations, each easing pressure on global fisheries.
• Meat and meat products may have a variety of fatty acid compositions depending on the species from which the tissue is derived, the type of tissue analyzed, and the feed provided to the animal.
• meat and meat products are usually characterized by an excess of saturated fatty acids and a dearth of beneficial fatty acids such as SDA, EPA, and DPA. As such, it would be preferably to increase the relative ratio of beneficial fatty acids to saturated fatty acids in meat products.
• references describing methods of altering the characteristics of meat products include US patents Nos.#. 6,955,830, 6,869,631, 6,763,760, 6,632,463, 6,405,646, 6,386,099, 6,319,527, 6,165,528, 5,071,666, 5,881,640, 5,307,737, 5,053,237, 3,556,809, 5,460,842, and 5,116,629, each of which is hereby incorporated by reference.
• additives may also be used to alter the quality of the product.
• Some meat products are processed to lower the animal fat content of the meat product. See for example US Patents Nos.# 6,099,891, 5,468,510, 5,382,444, 5,688,549, 5,460,842, 5,116,629 and PCT applications WO05034652A1, WO05094617A1 each of which is hereby incorporated by reference.
• Some methods of improving the fat content of meat products include heating the product to remove the solid-phase animal fat.
• US Patent 5,116,633 describes a process of heating a meat product in an unsaturated oil to extract the natural fats and increase the content of unsaturated fat and cholesterol.
• US Patent 5,460,842 describes a process for altering the fat content of meat by extracting fat using a cooled needle, and injecting "marbleizing fat” content alteration of raw meat useful for removing fat - by piercing raw meat with needle, subsequently withdrawing and removing adhered fat, useful for exchanging saturated fat for unsaturated.
• Modification of animal flesh in processing may include muscle tissue from bovine, swine, chicken, fish, or other sources. One particular example can be found in the pork industry.
• the pork industry has emphasized reducing the fat content in the finished product through methods of phenotypic breeding. More recently, marker assisted selection and other methods of advanced breeding have been used to create animals with altered meat quality characteristics.
• the long term effect of lowering the fat content of pig tissue through breeding is considered to be a healthier, lower fat meat product for the consumer.
• the lower fat products generally come at the expense negative impacts to of flavor and texture. It would be desirable to have both the health benefits of lower saturated fats, and the improved flavor and texture associated with higher fat content in the same product.
• Methods of marker assisted selection and markers associated with meat quality are known in the art. Examples include the following: US6803190, US20040261138A1, US20060288433Al, US20040018511Al, US6569629, US6803190, US6919177, US6458531, US6492142, US6919177, US7074562, and US20030186299Al, each of which is hereby incorporated by reference. Genetic markers can be used to more effectively select animals for breeding or slaughter based on desired phenotypes. In particular, marker assisted selection is more effective than traditional breeding methods when selecting for traits which are difficult to measure on a live animal such as intramuscular fat, meat quality, meat tenderness, and meat color.
• Embodiments of the present invention encompass meat products with increased levels of beneficial fatty acids. Preferable embodiments including stearidonic acid (18:4 ⁇ 3). Alternative, embodiments of the invention may improve the palatability of the meat product and/or the health of an end consumer through incorporation of these beneficial fatty acids. In particular embodiments of the invention, oil comprising stearidonic acid from transgenic soybeans is incorporated into the meat product.
• SDA oil provides an initial clean flavor, longer shelf-life stability and enhanced nutritional quality relative to traditional omega-3 alternatives.
• Embodiments of this invention may comprise a meat product with improved characteristics and improved compositions of physiologically significant polyunsaturated fatty acids.
• methods are disclosed for incorporating the SDA oil into a meat product. These methods of incorporation may include injection, physical mixing, surface contact, or any other methods known in the art. [0024] Furthermore, methods of making meat products comprising SDA oil and being relatively low in saturated fat are disclosed. These methods of making a meat product incorporate breeding techniques to alter the composition of the meat product, such as for example, using marker assisted selection to decrease the inter-muscular fat of the pig. These methods further comprise adding SDA oil to the meat product to increase the beneficial lipid content, thereby improving the taste and palatability of the product while improving the health profile of the resulting meat product.
• Methods for making a meat product may also comprise lowering the saturated fat content after tissue has been harvested from the livestock animal through mechanical or chemical means, along with the additions of SDA oil to increase the beneficial fatty acid content of the meat.
• Methods for making a meat product may also comprise processing the meat product at reduced temperatures and/or addition of SDA after high temperature processing steps.
• meat products according to compositions and/or methods described herein are incorporated into food products having substantial secondary components, such as for example, a pizza, a pre-packaged meal, soup, ready-to-eat meals, or other food product.
• meat color refers to measurements of color of meat obtained from a slaughtered animal.
• meat quality refers to measurements of the palatability and/or eating qualities (e.g. marbling, texture, and tenderness) of meat from a slaughtered animal.
• quantifying meat quality can be accomplished by measuring phenotypes including, but not limited to: Drip loss (%); purge loss (%); pH of loin or ham; moisture content in the loin (%); NPPC firmness quality score (1-5); Werner-Bratzler shear force (kg); intramuscular fat content (%), and cooking loss (%).
• the term "meat composition” refers to any number of phenotypes including, but not limited to: average backfat thickness (e.g. in inches or centimeters) measured on the carcass or on the live animal using ultrasound; loin muscle (eye) area (e.g. in 2 ) measured on the carcass or on the live animal using ultrasound; fat depth, loin depth
• carcass weight measured on-line after slaughter using industry standard equipment (e.g., Fat-O-Meater); predicted lean percentage (%) calculated from backfat thickness, loin muscle depth or area, and body or carcass weight; carcass length (e.g. in inches); National Pork Producers Council (NPPC) marbling quality score (i.e., 1-5);
• industry standard equipment e.g., Fat-O-Meater
• predicted lean percentage % calculated from backfat thickness, loin muscle depth or area, and body or carcass weight
• carcass length e.g. in inches
• National Pork Producers Council National Pork Producers Council
• Intramuscular fat content of the loin (%); primal cuts (loin, shoulder, belly, etc) as a percent of carcass weight (%); carcass yield as a percentage of live weight (%).
• meal product refers to food products comprising the flesh of livestock animals such as bovine, swine, and poultry.
• genetic marker refers to a DNA sequence that can be identified due to unique features such as a single nucleotide polymorphism (SNP), microsatellite, RFLP markers, or any other identifiable genetic marker.
• SNP single nucleotide polymorphism
• RFLP markers RFLP markers
• mixed meat product refers to products which incorporate multiple muscles into the same meat product through the process of mechanical mixing.
• Such products include, for example, sausages, hot dogs, hamburger, processed meats, bologna, salami, reconstituted meat products, and the like.
• Mixed meat products may include meat only from one species of animal or from more than one species of animal.
• Mixed meat products may also include protein from non-meat sources such as, for example, soy protein.
• the term "reconstituted meat product” includes meat products comprising a plurality of meat pieces agglomerated together.
• Reconstituted meat products may include additional components such as binders, fillers, flavorants, and preservatives, including additional sources of protein such as other meat from other sources, including plants or plant products for example - soy protein.
• the term "food product” refers to edible products for human consumption.
• Embodiments of the invention include edible meat products for human consumption comprising stearidonic acid (SDA), and gamma linolenic acid (GLA), wherein: the SDA concentration is at least about 0.01% of the total fat content of the meat product; the GLA concentration is at least about 0.01% of the total fat content of the meat product; and wherein the ratio of SDA/GLA concentrations is at least about 2.
• Preferred embodiments may have an SDA concentration which is at least about 0.1%, at least about 0.5%, or at least about 1.0% of the total fatty acid content of the meat product.
• Alternative embodiments may have a ratio of SDA/GLA concentrations which are at least about 2.0.
• Alternative embodiments may further comprise ALA and may have a ratio of SDA/ ALA concentrations which is at least about 1.0.
• Embodiments of this invention may comprise endogenous oil from a transgenic plant, preferably transgenic soybean oil.
• Embodiments of the invention may also comprise tocopherol, preferably at least 50ppm, and more preferably at least about lOOppm tocopherol. Preferably, embodiments have an omega-3 to omega-6 fatty acid ratio of the meat product which is greater than about 1:1.
• Alternative embodiments of the invention may further comprise 6-cis, 9-cis, 12-cis, 15- trans-octadecatetraenoic acid.
• Alternative embodiments of the invention may further comprise 9-cis, 12-cis, 15-trans-alpha linolenic acid.
• Alternative embodiments of the invention may further comprise 6, 9 -octadecadienoic acid.
• Embodiments of the invention may also comprise soy protein.
• Embodiments of the invention include meat products in a variety of forms including a pumped meat product, a mechanically mixed meat product, a reconstituted meat product, and a coated meat product.
• Embodiments of the invention may further comprise any additives known in the art as flavorings, tenderizing agents, stabilizers, emulsifiers, antioxidants, and/or processing aids for use with meat products.
• Additives may include, but are not limited to, sodium chloride, potassium chloride, tocopherol, sugar, sodium tripolyphosphate, ascorbate, sodium nitrite, sodium nitrate, potassium lactate, sodium diacetate, acetic acid, sodium ascorbate, smoke flavoring agents, lecithin, water, methyl cellulose, bouillon, vinegar, colorants, vitamins, preservatives, pickling agents, spices, binders, soy protein, and/or emulsifying agents.
• embodiments of the invention include food products made with meat products as described herein. Such food products may include any edible composition comprising a meat product, including such foods as pizza, soups, ready to eat meals, prepared dishes, and the like.
• Embodiments of the present invention also include a methods of producing a meat product comprising: testing an animal to obtain genetic information; selecting said animal for breeding according to said genetic information; breeding said animal to produce progeny; harvesting meat tissue from at least one of said progeny; contacting said meat tissue with a vegetable oil; and wherein said vegetable oil comprises at least about 0.01% stearidonic acid.
• the animal is selected based on predicted meat quality, intra-muscular fat, or pH, meat color, or meat composition.
• the animal is selected based on predicted intra-muscular fat content.
• said selection results in meat with a reduced fat content.
• Embodiments of the present invention also include methods of producing a food product for human consumption comprising: providing a source of stearidonic acid; providing an aqueous liquid; providing at least one meat ingredient; contacting said stearidonic acid source with said aqueous liquid and said meat ingredient to make a supplemented meat product; and wherein said stearidonic acid source comprises a transgenic plant oil.
• Embodiments of the present invention also include methods of making a food product for human consumption comprising: providing a source of stearidonic acid (SDA); providing animal meat tissue; heating said animal meat tissue; and combining said source of SDA with said animal tissue.
• Preferred embodiments of this invention include methods of making a food product wherein said meat tissue is heated prior to combining with said source of SDA.
• Alternative embodiments include heating said meat products to remove a portion of the animal fat prior to combining with said source of SDA.
• Preferable embodiments include minimizing the temperature used to process said meat after combining with said SDA source.
• Embodiments of the present invention also include food products comprising arachidonic acid (AA) and stearidonic acid (SDA) wherein the ratio of concentrations of SDA/ AA is at least about 0.1, and wherein said food product comprises at least about 0.1 grams of SDA per 100 grams of food product.
• AA arachidonic acid
• SDA stearidonic acid
• Preferable embodiments of the invention comprise ratio of concentrations of SDA/ AA is at least about 0.2, at least about 0.3, at least about 0.5, at least about 1.0, and at least about 2.0.
• Embodiments of the present invention may further comprise gamma-linolenic acid (GLA).
• GLA gamma-linolenic acid
• embodiments of the present invention may comprise a ratio of SDA/GLA of at least about 2.0.
• Embodiments of the present invention also comprise methods of producing meat products comprising: providing animal tissue; reducing the saturated fatty acid content of said animal tissue; contacting vegetable oil with said animal tissue; wherein the unsaturated fatty acid content of said meat product is increased and wherein said meat product comprises at least about 0.1% SDA.
• Embodiments of the invention may further comprise heating said animal tissue while producing said meat products.
• the vegetable oil comprises an endogenous soybean oil.
• the vegetable oil further comprises gamma-linolenic acid (GLA).
• GLA gamma-linolenic acid
• the vegetable oil has a ratio of SDA/GLA of at least about 2.0.
• Alternative embodiments of the invention may further comprise arachidonic acid (AA).
• AA arachidonic acid
• Preferable embodiments may have an SDA/ AA concentration ratio of at least about 0.1, at least about 0.2, at least about 0.3, at least about 0.4, at least about 0.5, at least about 1.0, and at least about 2.0.
• Methods of making embodiments of the invention may comprise physically mixing the vegetable oil and the meat tissue, injecting said vegetable oil into said meat tissue, or contacting exterior surfaces of said meat tissue with vegetable oil. Production of SDA:
• Embodiments of the present invention relate to a system for an improved method for the plant based production of stearidonic acid and its incorporation into the diets of humans and livestock in an effort to improve human health.
• This production is made possible through the utilization of transgenic plants engineered to produce SDA in sufficiently high yield to so as to allow commercial incorporation into food products.
• the acid and salt forms of fatty acids for instance, butyric acid and butyrate, arachidonic acid and arachidonate, will be considered interchangeable chemical forms.
• oils from transgenic plants have been created.
• Some embodiments of the present invention may incorporate products of transgenic plants such as transgenic soybean oil.
• Transgenic plants and methods for creating such transgenic plants can be found in the literature. See for example, WO2005/021761A1.
• Table 2 the composition of the transgenic soy oil is substantially different than that of the accepted standards for soy oil.
• the SDA rich soybeans produced in a recombinant oilseed plant provides a composition not previously available for food product manufacturers.
• the use of these compositions is made possible without the traditional concerns with stability when oils comprising DHA are delivered from a fish or algal source.
• the preferred source of stearidonic acid in some embodiments is transgenic soybeans which have been engineered to produce high levels of stearidonic acid.
• the soybeans may be processed at an oil processing facility and oil may be extracted consistent with the methods described in US Patent Applications 2006/0111578A1, 2006/0110521 Al, and 2006/0111254A1, herein incorporated by reference.
• Fatty acid compositions of various meat products are known in the art. See for example Droulez et al (2006), Rule et al (2002), and the USDA Composition of Foods - Raw, Processed, Prepared; USDA Human Nutrition Information Service Agriculture Handbook. Fatty acid compositions for numerous meat products are also available on the internet. See for example, the USDA Agricultural Research Service Nutrient Data Laboratory at www.nal.usda.gov/fnic/foodcomp/search/. For exemplary purposes, Table 3 contains fatty acid profiles excerpted from Rule et al (Comparison of muscle fatty acid profiles and cholesterol concentrations of bison, beef cattle, elk and chicken, J Anim Sci.
• the present invention include meat and meat products comprising both animal tissue and steardidonic acid.
• addition of stearidonic acid can improve the amount of beneficial fatty acids in proportion to less desirable fatty acids such as omega-6 fatty acids like arachidonic acid (AA).
• the ratio of concentrations of SDA to arachidonic acid (20:4n-6) is at least about 0.1, at least about 0.2, at least about 0.3, at least about at least about 0.5, at least about 1.0, and at least about 2.0.
• Genomic technology offers the potential for greater improvement in traits of livestock production and meat quality through the discovery of genes, or genetic markers linked to genes, that account for genetic variation and can be used for more direct and accurate selection. Numerous markers with associations with meat production and quality traits have been reported (see Rothschild and Plastow, 1999; see in particular pp 1-8 ).
• pigs harvested for pork around the world are typically produced by crossing closed breeding populations with distinct characteristics for maternal productivity, growth efficiency and meat quality. These closed populations typically have low levels of inbreeding and genome scans yielding markers that can be used across the population for selection. These resulting linked markers can be used in several methods of marker or marker-assisted selection, including whole-genome selection (Meu Giveaway et al., 2001) to improve the genetic merit of the population for these traits and the value created in the pork production chain.
• Some embodiments of the invention provide methods for evaluating an animal's genotype at one or more positions in the animal's genome for MAS prior to further processing.
• the animal's genotype is evaluated at a position within a segment of DNA (an allele) that contains at least one genetic marker such as, for example a single-nucleotide polymorphism (SNP) or a micro satellite marker.
• SNP single-nucleotide polymorphism
• the genomic sequence at the marker locus may be determined by any means compatible with embodiments of the present invention.
• Phenotypic traits that may be associated with the SNPs include, but are not limited to: growth traits, body and/or carcass composition, meat quality, intra-muscular fat, pH, and meat color.
• MAS is used to alter these phenotypic traits to more effectively address productivity and product quality needs for products comprising added beneficial fatty acids such as stearidonic acid.
• animals identified as having SNP alleles associated with desirable phenotypes are allocated for use consistent with that phenotype (e.g. allocated for breeding based on phenotypes positively associated with improved growth).
• animals that do not have SNP genotypes that are positively correlated with the desired phenotype (or possess SNP alleles that are negatively correlated with that phenotype) are not allocated for the same use as those with a positive correlation for the trait.
• determination of whether and how an animal will be used as a potential parent animal may be based on its genotype at one or more, 10 or more, 25 or more, 50 or more, or 100 or more genetic markers.
• Other aspects of these embodiments provide methods where analysis of one or more SNPs is combined with any other desired genomic or phenotypic analysis (e.g. analysis of any genetic markers beyond those disclosed in the instant invention).
• the SNP(s) analyzed may all be selected from those associated only with growth, or only with composition, or only meat color, or only meat quality. Conversely, the analysis may be done for SNPs selected from any desired combination of these or other traits.
• animals based on the composition of the meat, such as for example intramuscular fat (IMF) content or total fat content.
• IMF intramuscular fat
• animals are selected for breeding based on markers which are associated with relatively lower IMF content.
• the meat from these animals are further modified after slaughter by incorporation of oils comprising beneficial fatty acids such as stearidonic acid, thereby raising the total lipid content of the finished product while maintaining dual health benefits of low- saturated fats and high SDA content in a meat product.
• Quantifying meat IMF % is generally known in the art. Differences exist in measurement methods, particularly across livestock species. However, one of skill in the art can readily ascertain methods for quantifying the relevant phenotypic parameters. For example, in pigs measuring IMG can be accomplished by measuring phenotypes including, but not limited to: National Pork Producers Council (NPPC) marbling standard guidelines (1.0-10.0) at the 10 th /! 1 th rib intersection of the longissimus dorsi (loin); or Total Lipid Percent can be determined through chemical fat extraction of a emulsified loin sample collected near the 10 th /! 1 th rib intersection of the longissimus dorsi (Folch, J., M.
• NPPC National Pork Producers Council
• one or more of the markers with significant association to that trait can be used in selection of breeding animals.
• use of animals possessing a marker allele (or a haplotype of multiple marker alleles) in population- wide LD with a favorable QTL allele will increase the breeding value of animals used in breeding, increase the frequency of that QTL allele in the population over time and thereby increase the average genetic merit of the population for that trait. This increased genetic merit can be disseminated to commercial populations for full realization of value.
• a closed genetic nucleus (GN) population of pigs may be maintained to produce boars for use in commercial production of market hogs.
• a GN herd of 200 sows would be expected to produce approximately 4,000 offspring per year (2,000 boar offspring per year), typically according to a weekly breeding and farrowing schedule.
• a commercial sow herd of 300,000 would be expected to produce more than 6,000,000 market pigs per year and might maintain a boar stud with inventory of 1,500 boars to service the 300,000 sows.
• the GN herd of 200 sows could support boar replacements for the 300,000 sows of commercial production by shipping 500 boars every six months to the commercial boar stud (assumes inventory in the commercial boar stud is turned over every 1.5 years and the top half of boar candidates from the GN are selected for commercial breeding).
• the first step in using a SNP for estimation of breeding value and selection in the GN is collection of DNA from all offspring that will be candidates for selection as breeders in the GN or as breeders in other commercial populations (in the present example, the 4,000 offspring produced in the GN each year).
• One method is to capture shortly after farrowing a small bit of tail tissue from each piglet into a labeled (bar-coded) tube.
• the DNA extracted from this tissue can be used to assay an essentially unlimited number of SNP markers and the results can be included in selection decisions before the animal reaches breeding age.
• One method for incorporating into selection decisions the markers (or marker haplotypes) determined to be in population- wide LD with valuable QTL alleles is based on classical quantitative genetics and selection index theory (Falconer and Mackay, 1996; Dekkers and Chakraborty, 2001).
• a random sample of animals with phenotypic measurements for the trait of interest can be analyzed with a mixed animal model with the marker fitted as a fixed effect or as a covariate (regression of phenotype on number of allele copies). Results from either method of fitting marker effects can be used to derive the allele substitution effects, and in turn the breeding value of the marker:
• (X 1 and ⁇ 2 are the average effects of alleles 1 and 2, respectively; ⁇ is the average effect of allele substitution; p and q are the frequencies in the population of alleles 1 and 2, respectively; a and d are additive and dominance effects, respectively; g A i A i, g A i A2 and g A2A2 are the (marker) breeding values for animals with marker genotypes AlAl, A1A2 and A2A2, respectively.
• the total trait breeding value for an animal is the sum of breeding values for each marker (or haplotype) considered and the residual polygenic breeding value:
• EBV y is the Estimated Trait Breeding Value for the i ⁇ animal
• n is the total number of markers (haplotypes) under consideration
• U 1 is the polygenic breeding value for the i th animal after fitting the marker genotype(s).
• meat products are made using meat from animals that have been selected using MAS.
• these animals are selected to modify the meat composition or quality.
• MAS is used to lower the intramuscular fat in swine prior to incorporation of SDA oil into the meat product.
• markers and associated methods which can be used for breeding related to meat quality, productivity, and/or meat composition traits for livestock animals such as swine, bovine, fish, crustaceans, and poultry include the following, herein incorporated by reference: US5944652, US6458531, US6696253, US6716972, US6803190, US6919177, US7244564, US2002051989A1, US2003017470A1, US2003129610A1, US2003186299A1, US2004018511A1, US2004029145A1, US2004048267A1, US2004055029A1, US2004235030A1, US2004259127A1, US2004261138A1, US2005202484A1, US2005208551A1, US2005214814A1, US2006037090A1, US2006211006A1, US2006223058A1, US2006275793A1, US2006288433A1, US2007003956A1, US2007006333A1, US2007092909A1, WO0006777
• Examples of meat processing methods include the following US Patents Nos: and applications, each of which is herein incorporated by reference: US3556809, US3916777, US4463027, US4584204, US4778682, US4867986, US4904496, US4980185, US5053237, US5082678, US5106639, US5116629, US5116633, US5211976, US5213829, US5250006, US5256433, US5380545, US5382444, US5415883, US5460842, US5468510, US5472725, US5474790, US5484625, US5489443, US5492711, US5514396, US5523102, US5556662, US5631035, US5674550, US5688549, US5698255, US5807598, US5895674, US5965191 , US5989601, US6014926, US6054147, US6099891, US6103276, US6248381, US66
• the stearidonic acid will be incorporated into the meat product in the form of a liquid vegetable oil, such as for example in a transgenic soybean oil.
• This fluid may be injected into the meat, mechanically mixed with the meat, or used to coat the exterior of the meat.
• the oil may be first emulsified for form an emulsion.
• the emulsion comprises an oil containing SDA and an aqueous liquid.
• the amount of fat in the starting meat is altered through sophisticated breeding methods such as marker assisted selection.
• some embodiments of the invention may also incorporate alternative means of reducing the fat content of meat such as pre-cooking some portion of the meat to reduce the fat content, incorporating meat from species known to have lower fat content, such as chicken breast meat for example, and any other extraction techniques such as are known in the art.
• a variety of injection mechanisms are known in the art which may be suitable for application in embodiments of the present invention.
• some devices utilize a plurality of needles inserted into the meat. As the needles are removed, a fluid is injected into the space formed by the injection needle.
• Alternative methods employ a plurality of nozzles with a high pressure fluid (e.g., injectate) which is forced into the meat using the pressure differential.
• the injectate may be heated in a water bath at a temperature of from about 110° F to about 160° F to keep the injectate within an acceptable temperature range.
• Meat can be placed at varying distances (from as close as 0.1 inch to as far as 10 inches) below the injector nozzles.
• Streams of liquid injectate may be emitted from the nozzles at high velocity, with bursts lasting 0.1 to 10 seconds, preferably 0.5 to 5 seconds..
• the liquid is pressurized to from about 200 pounds per square inch ("psi") to about 8,500 psi. to obtain a high velocity injectate stream.
• the meat is preferably injected at multiple sites. The distance between injection sites was variable in order to achieve particular tenderization, moisture, and fat percentage characteristics.
• the injectate may be comprised of various oils, water, and binders, is preferably maintained at below 120° F. in order to avoid precooking of the meat and to minimize any degradation of oils (e.g., SDA) in the injectate. More preferably, the injectate is maintained at below 100° F. In addition to incorporation of the fluid, mechanical tenderization may also result from the injection process.
• oils e.g., SDA
• mechanical tenderization may also result from the injection process.
• the spatial distribution of the SDA oil is predetermined.
• he energy of the injectate stream is dissipated along the path of the injectate.
• injectate may be relatively uniformly distributed throughout the meat.
• this procedure results in distribution of oils, water, and flavoring through the interior of the meat tissue.
• a predetermined spatial orientation may provide an excellent technique for injecting oils, emulsions, or even an enzyme, such as collagenase, which is known to break down connected tissue.
• injecting collagenase will tend to result in rapid and efficient tenderization since the injectate follows the tough connective tissue throughout the meat. The collagenase is, therefore, located directly adjacent to the tissue on which it acts.
• binders are added to the injectate stream.
• Such binders may include gel-forming materials and emulsifiers in the amount of from about 0.1% to about 2.5% of the total injectate.
• Binders may include gums, thickeners, salts, and combinations of the same.
• Optional ingredients incorporated into or on the meat may further comprise a variety of additional compositions such as sodium chloride, potassium chloride, tocopherol, sugar, sodium tripolyphosphate, ascorbate, sodium nitrite, sodium nitrate, potassium lactate, sodium diacetate, acetic acid, sodium ascorbate, smoke flavoring agent, lecithin, water, methyl cellulose, bouillon, vinegar, colorants, vitamins, preservatives, pickling agents, spices, binders, and/or emulsifying agents.
• additional compositions such as sodium chloride, potassium chloride, tocopherol, sugar, sodium tripolyphosphate, ascorbate, sodium nitrite, sodium nitrate, potassium lactate, sodium diacetate, acetic acid, sodium ascorbate, smoke flavoring agent, lecithin, water, methyl cellulose, bouillon, vinegar, colorants, vitamins, preservatives, pickling agents, spices, binders, and/or emulsifying agents.
• stabilizers and antioxidants may be used to maintain the flavor and organoleptic properties of both the meat and oil.
• tocochromanols such as tocopherol can be incorporated into the oil and/or meat product.
• Additional stabilizers may include tertiary butyl hydroquinone (TBHQ), butylated hydroxyamisole, butylated hydroxytoluene, and citric acid.
• the product may be sold without additives, or additives may be used to enhance the flavor, texture, and/or appearance of the product such as by injection of a brine solution or coating with a marinade.
• Methods of injecting solutions into meat include piercing the meat with needles and injecting a fluid under pressure into the interior of the meat (for example, US Patents Nos. #: 5,071,666, 5,142,971, 5,176,071, 6,165,528, and 6,497,176, herein incorporated by reference).
• a soybean oil comprising stearidonic acid is injected into the meat product.
• the SDA- containing oil may further comprise traditional brine additives.
• the SDA- containing oil may further comprise an emulsifier and be injected in the form of an emulsion.
• meat sources and additives may be used in the same product. Meat from various parts of an animal may be combined including various meat sources such as meat trimmings, mechanically separated meat, meat obtained through recovery techniques, and/or meat analogs.
• Alternative sources of protein may be added such as meat from other animal species or even plant-based protein such as soy protein (see for example, US patent 6,797,288, herein incorporated by reference).
• extenders, fillers or binders may be used particularly for reformed meat products such as for example ground meats and/or reconstituted meats processed into cold-cuts.
• Such fillers may include soy flour, soy protein concentrate, isolated soy protein, wheat, rice, corn, barley, vegetable starch, vegetable flour, non-fat dry milk, skim milk, and dried milk.
• Extenders, fillers and binders typically comprise less than about 4% of the total meat product.
• mechanically mixed products may be formed into secondary shapes such as sausages, loafs, patties, links, cylinders, or other forms. This process may involve extrusion, injection into a casing, or molding under pressure.
• the meat product may be heated and/or at least partially cooked prior to sale to the consumer.
• the meat product may be heated and/or at least partially cooked prior to the addition of the SDA comprising composition, such as transgenic soy oil comprising SDA.
• at least some portion of the meat product is substantially cooked prior to addition of SDA.
• the complete SDA containing meat product is not exposed to high temperatures for long periods of time prior to delivery to the consumer. In some cases, high temperature may be acceptable for short periods of time.
• meat products according to compositions and/or methods described herein are incorporated into food products having substantial secondary components, such as for example, pizza, prepared dishes, frozen dinners, soups, sandwiches, hamburgers, ready to eat meals, and the like.
• the two patties were placed in a frying pan and the pan was placed on a household stove. The patties were fried until done. Each patty was rotated to avoid burning of the meat. The patties were removed from the heat when done (based on visual inspection) and cooled down using ambient air. The patties were packaged in a container for subsequent sensory and analytical testing.
• a control was produced in the same manner as above, but with commercial soy oil not containing SDA. Sensory evaluation of the control and SDA- containing hamburgers indicated no significant changes to texture or flavor.
• a whole muscle or portion of a whole muscle of pork is coated in a liquid mixture at 10% of less of the precooked muscle weight.
• the marinade solution should be composed of the following ingredients listed as a percent by weight included in marinade solution: 66% SDA soy oil, 26% wine (white), 5% sage, and 3% black pepper. Lemon juice and minced garlic can be added to marinade solution by taste.
• the marinade solution should be coated over the pre-cooked whole muscle cut and refrigerated for 8-12 hours prior to cooking. This marinade solution could also be used by food processors prior to packaging.
• Example 3 Preparation of a Turkey meat product comprising an SDA emulsion.
• a portion of whole, single muscle meat derived from turkey is injected with an emulsion of transgenic SDA-containing soybean oil (15% SDA) and water, also containing salt, sugar and emulsifier, in an amount approximately 15% by weight of meat.
• the body was then tenderized, and subjected to a tumbling operation at 6 rpm for a period of 40 minutes at a pressure of 1.5x104 Pa.
• the portion is then placed in a tray, and conformed to a generally rectangular shape and frozen to a temperature of minus 18° C. into a condition in which the portion is mechanically rigid.
• the block produced by freezing the portions may then be removed from the tray, and bagged and retained frozen until desired for further use.
• the partially- formed portion thus provided is placed in a high pressure mould in which it is formed into a final desired shape, while retaining its low temperature, at a high pressure of between 500 and 10,000 p.s.L, preferably between 1,000 and 5,000 p.s.L, for a period of about 7.5 seconds. While the high pressure is applied to the portion, the meat product conforms to the shape of the mould without any significant disruption of the muscle tissue.
• the block On conclusion of the pressing operation the block is removed from the high pressure mould, and is then subjected to a slicing operation, during which it is sliced to produce portions of meat having a relatively uniform cross section and having a thickness determined by the thickness of the cut.
• the slicing operation may be carried out at a temperature raised somewhat about the -18° C. at which the moulding operation takes place, but preferably slicing is carried out without any significant elevation in temperature.
• the slices may then be vacuum sealed in individual bags, and retained chilled or frozen until the product is cooked by the consumer.
• the meat may be cooked directly after being sliced, whilst in an individual pack, such as in a continuous oven or in a water bath, and frozen again or chilled, being reheated prior to being consumed.
• the meat may be cooked prior to the addition of the SDA to minimize any degradation of the added oil.
• Example 4 Turkey Meat with SDA Oil.
• Portions of whole muscle turkey meat are first injected with a marinade solution to approximately 12% of their original weight, with the brine composition comprising 78.5% water, 6% salt, 7% transgenic soy oil, 7% turkey flavor (salt, wheat starch, chicken fat, beef extract and alpha tocopherol), and 1.5% sodium caseinate, by weight.
• the SDA containing soy oil has an SDA concentration of about 28%.
• An injector equipped with bayonet-typed tenderizing needles are used to effect tenderizing of the meat.
• the injected/tenderized portions are tumbled under a low pressure of 1.5x104 Pa or lower for 5 hours at 6 rpm.
• the tumbled portions are separated and graded into various weight categories to accommodate various sizes of moulds.
• the graded portions are fitted into the moulds in close proximity to fill the moulds, with care being taken to ensure alignment of the grain of the portions.
• a plurality of filled mould assemblies are placed into a plate-freezer 23, and subjected to a relatively low pressure of between 1 and 5 p.s.i. to conform the portions to the shape of the compartments without causing any significant disruption of the muscle tissue and are reduced in temperature by the plate-freezer to a temperature of about -18° C over a period of about 4 hours.
• Example 2 The partially-formed portions of Example 2 are placed in a high pressure mould
• FIG. 6 in which they are formed into a final desired shape, while being retained at its low temperature and at a pressure of between 500 and 10,000 p.s.i. (3.45x106 to 6.9x107 Pa) preferably about 3,000 p.s.i., for a period of about 7.5 seconds.
• the block On conclusion of the pressing operation the block is removed from the high pressure mould, and is then subjected to a slicing operation, during which it is sliced to produce portions of meat having a relatively uniform cross section and having a thickness determined by the thickness of the cut.
• the slicing operation may be carried out at a temperature raised somewhat about the -18° C. at which the molding operation takes place, but preferably slicing is carried out without any significant elevation in temperature.
• the slices may then be vacuum sealed in individual bags, and retained chilled or frozen until the product is cooked by the consumer.
• the meat may be cooked directly after being sliced, whilst in an individual pack, such as in a continuous oven or in a water bath, and frozen again or chilled, being reheated prior to being consumed.
• the meat may be cooked prior to the addition of the SDA to minimize any degradation of the added oil.
• Example 5 Beef product comprising SDA Oil
• Top loin and chuck portions of beef are tenderized and subsequently injected with a brine solution.
• the brine composition was 81.5 wt % water, 6% salt, 3% dextrose, 1.5% sodium caseinate, and 8% soy oil.
• the roasts are then tumbled for 40 minutes at 6 rpm under 26" Hg vacuum.
• the portions are then placed in a mould and formed generally to the shape of the mould under a relatively low pressure of 5 p.s.L, and while in the mould are subjected to a temperature of -15° C. until they are rigid throughout.
• the portions While rigidly frozen, the portions were transferred to a high pressure mould and pressed for seven seconds at 1000 p.s.L, the portions being conformed to the shape of the high pressure mould. The portions were then sliced into steaks to a thickness of 0.69 inches (15 mm).
• a processed pork meat products of can be prepared having the following formulation: raw pork trimmings 93% (75% lean); Salt 1.5%; White pepper: 0.3%; Sage: 0.1%; Mace: 0.1%; Ginger: 0.02%; SDA containing soy oil 3%; High fructose corn syrup 1.5% (90 D. E.); wherein each percentage is on a weight basis.
• the soybean oil may also contain 100 ppm TBHQ; 1,000 ppm tocopherol, and 30 ppm citric acid.
• the SDA containing soy oil has an SDA concentration of about 28%.
• a small quantity of the above formulation is prepared by making a pre-blend of all dry ingredients and a second pre-blend of all wet ingredients.
• the dry pre-blend is admixed with the wet pre-blend in a mixer.
• the admixture is then added the oil based ingredients.
• the composition is further mixed at high shear to form a stable emulsion.
• the stable emulsion is then injected into sausage casings, packaged and stored.
• a similar process can be used to make a pre-cooked product wherein the pork trimmings are cooked prior to combination with the SDA containing soy oil.
• a skinless polish sausage is prepared according to the procedure of Example 1 having the following ingredients: Beef trimmings 45% (80% lean); Pork cheeks 20%; Regular pork trimmings: 25%; Salt: 2%; Ground black pepper: 0.15%; Marjoram: 0.05%; Sodium nitrite 0.02%; Garlic: 0.05%; SDA Soy Oil 6.5%; High fructose corn syrup 1.5%.
• the soybean oil may also contain 100 ppm TBHQ; 1,000 ppm tocopherol, and 30 ppm citric acid.
• the SDA containing soy oil has an SDA concentration of about 20%.
• a small quantity of the above formulation is prepared by making a pre-blend of all dry ingredients and a second pre-blend of all wet ingredients.
• the dry pre-blend is admixed with the wet pre-blend in a mixer.
• To the admixture is added the oil based ingredients.
• the composition is further mixed at high shear form a stable emulsion.
• the stable emulsion is then injected into sausage casings, packaged and stored.
• the following ingredients can be used to make a hard salami product containing SDA.
• the ingredients include: beef chunks: 40%; pork jowls - 40%; Regular pork trimmings: 20%; salt: 3.2%; sucrose: 1.5; white pepper: 0.2%; sodium nitrate - 0.1%; garlic powder: 0.1%; SDA soy oil 3%; antioxidant blend: 0.01%; high fructose corn syrup 1%.
• the SDA containing soy oil has an SDA concentration of about 20%.
• the antioxidant blend referred to above consists of 60% tocopherol available from Henkel Corporation under the trade name CoviOX T70, 30% TBHQ mixture of (32% glycerol monoleate, 30% corn oil, 20% TBHQ, 15% propylene glycol and 3% citric acid) available from Eastman Chemical Products Inc. under the trade name TENOX 20A, and 10% partially hydrogenated soybean oil with BHA to help protect flavor; available from Durkee Industrial Foods, under the trade name Durkex 100.
• the sausages are prepared by first grinding the beef through a 1/8-in. plate and the pork through a 1/4-in. plate. All the ingredients are then mixed in a mixer for 5 minutes or until a good distribution of the lean meat and fat is apparent.
• the mix is then stored in trays 8 to 10 inches deep for 2 to 4 days at 40° to 45° F. to age and to ferment the emulsion.
• the aged and fermented emulsion is next stuffed into No.
• the stuffed product is dried for 9 to 11 days at 40° F. and 60% relative humidity.
• Hot dogs are made with reduced fat pork processed.
• a continuous run of ground pork was comminuted through a Cozzini grinder, the exit temperature being 65° F.
• the resulting comminuted pork was passed through a swept surface heat exchanger, its temperature being raised to and maintained between 104° and 106° F. At this temperature, the flow of pork was fed into a decanter centrifuge.
• the fat reduced meat was mixed in a paddle or ribbon mixer with sodium nitrite, sodium erythorbate, salt and sodium phosphate.
• One quarter of the water needed for the hot dog formulation is added to a running mixer, and mixing proceeds for 10 minutes at a temperature of 30°-35° F.
• a curing salt solution is prepared by adding 90% water to a mixing vessel.
• the water is cooled to 2° F. and added in order are: 0.5% sodium tripolyphosphate, 5% Kilograms sodium chloride, 0.6% sodium nitrite, 5% Supro® Systems Ml 12, 3% ProliantTM 8610, 1% SDA containing soy oil, 1% dextrose, 4% maltodextrin, 1% carrageenan 0.2% sodium erythorbate, and 3% potato starch.
• a brine is formed having an SDA content.
• Hams are injected with this brine to give a meat product with improved fatty acid content.
• Example 11 Preparation of a Low- Saturated Fat Ham comprising SDA oil
• Swine from the Large White breed are bred using marker assisted selection of the MC4R gene to decrease the amount of fat produced in the pigs.
• the total fat content in the hams is reduced by approximately 1%.
• After slaughter, these hams are injected with an injectate comprising 2% SDA oil, but otherwise as described in Example 10 above.
• the resulting hams have a lower saturated saturated fat content, higher beneficial fatty acid content, and equivalent total fat content in comparison with hams produced using the method of Example 10.

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