AU2001258113B2 - Protein and lipid sources for use in aquafeeds and animal feeds and a process for their preparation - Google Patents

Description

-1- PROTEIN AND LIPID SOURCES FOR USE IN AQUAFEEDS AND ANIMAL FEEDS AND A PROCESS FOR THEIR PREPARATION Field of the Invention The present invention relates to a novel process for the production of nutritionally upgraded protein and lipid sources for use in aquafeeds and other animal feeds.

More specifically, the present invention relates to a process involving the coprocessing of animal offal(s) with oilseed(s); the invention also relates to products produced thereby.

In addition, the invention relates to cold pressed plant oils suitable for organic human foods, as well as products for use as components in organic fertilizers, both produced by the process of the invention.

In this specification, where a document, act or item of knowledge is referred to or discussed, this reference or discussion is not an admission that the document, act or item of knowledge or any combination thereof was at the priority date, publicly available, known to the public, part of the common general knowledge or known to be relevant to an attempt to solve any problem with which this specification is concerned.

Background of the Invention Feed accounts for on average 35-60% of the operating costs of salmon farms and it represents the largest cost in the culture of other carnivorous aquatic species.

Moreover, the protein sources presently used account for about 51% of the total feed cost and this percentage can be higher than this when increased reliance is placed on imported premium quality fish meals. The latter mainly originate from South America through the processing of whole pelagic fish species like sardines and anchovies and they are used to meet most of the dietary protein needs of farmed Canadian salmon. Accordingly, salmon farming profitability is marginal in Canada.

Currently, aquatic feeds contain high levels of fish meal and oil, which are mostly imported, to produce a protein-rich and sometimes lipid-rich salmon diets) aquatic feed. However, as noted hereinabove, such fish meal and oil can be very -2expensive and this will be especially true in the future due to progressively increasing demands that are being placed on the finite global supplies of fish meal and oil. Hence, altemative economical sources of protein and lipid are required.

One known approach is to use less expensive plant protein sources in aquafeed that have been specially processed so that they are in the form of nutritionally upgraded protein meals, concentrates, and isolates. These may be used either singly or in combination with rendered animal protein ingredients such as poultryby-product meal. To date, each of these protein products, such as canola meal, soybean meal, and poultry-by-product meal have been processed (produced) separately and then these protein sources have been blended together in dried and finely ground form in appropriate ratios for a particular aquatic species at the time of diet formulation and preparation.

U.S. Patent No. 4,418,086 to Marino et al. discloses the preparation of an animal feed which comprises a proteinaceous matrix, fat or oil, a sulfur source, farinaceous material, a plasticizer and water. The method disclosed involves the blending of the ingredients together introducing the mixture into an extruder and subjecting it to shear forces, mechanical work, heat and pressure such that the product temperature prior to discharge is at least 280 degrees F.

This patent is concerned with the production of an animal feed with a "meat like texture".

U.S. Patent No. 3,952,115 to Damico et al. relates to a feed where an amino acid is utlized as an additive to fortify a proteinaceous feed.

U.S. Patent No. 4,973,490 to Holmes discloses the production of animal feed products utilizing rape seed in combination with another plant species.

U.S. Patent No. 5,773,051 to Kim relates to a process for manufacturing a fish feed which refloats after initially sinking. This document discloses a process including blending conventional fish feed containing fish meal, wheat meal, soybean meal and other substances and compressing the mixture at a constant temperature to produce a molded product.

-3- Summary of the Invention There is provided in one embodiment of the present invention a process for the preparation of human grade oil, nutritionally upgraded oil seed meals, protein concentrates, lipid rich meals having a reduced fibre content and animal feed grade oils for use in fish or other non-human animal diets comprising providing a source of oilseed; pretreating the oilseed by one of: heat treating sufficiently to substantially reduce the concentration of at least some heat sensitive enzymes or antinutritional components normally present in the oilseed to obtain heat-treated seed that is comminuted by cold pressing to obtain human-grade oil or grinding; or, cold pressing the oilseed under conditions to substantially reduce particle size of the oilseed and obtain pressed raw seeds; and, blending the pretreated oilseed with unhydrolyzed comminuted or chopped animal protein sources together with an antioxidant, and when required to facilitate cooking and the extensive removal of water soluble antinutritional factors and carbohydrates in the oilseed, including water to produce a mixture thereof; cooking the mixture under conditions to substantially improve protein digestibility, and substantially free cellular water present in the animal protein source and facilitate separation of protein from the lipid in the animal protein source and the oilseed to obtain a cooked mixture; and, separating the cooked mixture into a stickwater fraction, a mostly insoluble protein-rich fraction, and an animal feed grade oil fraction.

In a further embodiment of the above, it is preferable the pretreating comprises a drying step, by cold pressing or by heat treating, and the dried heat treated seed is dehulled to produce a meat fraction and a hull fraction with further cold pressing being carried out on the meat fraction sufficiently to yield a high value human 18-12-05; 9:04AM;MSJ Mel 2 9296 399G 2/ 2 O -4c grade oil which is separated and a protein-and-lipid-rich meal, and blending this resulting Smeal with the protein source before cooking.

Desirably, in the above embodiments, the pretreating and blending steps include:

NO

drying the seed to reduce its moisture content to below about 10% by weight to obtain a dried seed or subjecting the oilseed to heat treatment under conditions Sselected to substantially deactivate, destroy or reduce the concentration of at least some of the antinutritional components normally present in oilseed to produce a 0 0 heat-treated seed; S- blending the oilseed and the animal protein source in a weight ratio of about o 10 10:90 to about 90:10, to form a mixture thereof.

In another embodiment of the present invention there is a provided a blended cooked feed product comprising: an oilseed-derived component selected from at least one of cooking water solubilized material, protein-rich insolubles and feed grade oil wherein said cooking water solubilized material, protein-rich insolubles and feed grade oil are produced according to the process of the Invention; and, an unhydrolyzed animal protein-source-derived component selected from at least one of cooking-water-solublized material, protein-rich insolubles and feed grade oil wherein said cooking water solubilized material, proteinrich insolubles and feed grade oil are produced according to the process of the invention.

Preferably, the cooked product comprises an animal feed grade oil comprising a cooked admixture of lipids from at least one of an oilseed and animal protein source.

It is further preferable the product comprises a protein concentrate suitable for use in fish and non-human animal diets, the oilseed comprising a heat-treated dehulled oilseed and the protein concentrate having: from about 38% to about 58% protein on a dry weight basis; from about 52% to about 77% protein on a lipid-free dry weight basis; from about 2.7% to 4.6% methionine and cystine calculated as a percent of protein; COMS ID No: SBMI-02213318 Received by IP Australia: Time 09:07 Date 2005-12-16 from about 4.3% to about 7.9% lysine calculated as a percent of said protein; from about 24% to 37% lipid on a dry weight basis; from about 1.7% to 10% crude fibre on a lipid-free dry weight basis reduced concentrations of water soluble antinutritional factors in an amount of at least 20% by weight relative to non-treated oilseed protein.

Moreover, the feed, having from about 40 to about 80% protein calculated on a lipid-free dry weight basis, being adapted for use in animal and aquafeeds whereby the admixture is characterized by at least one of the following: enriched concentrations of essential amino acids and bio-available minerals relative to those present in said animal offal or untreated oilseed; enriched concentrations of highly unsaturated n-3 fatty acids relative to those present initially in said oilseed if the source of animal offal is fish; reduced concentrations of heat-labile and water soluble and antinutritional factors in an amount of at least 20% by weight relative to non-treated oilseed protein; increased protein digestibility relative to non-treated oilseed protein; and a lipid concentration of less than 12% of dry weight of the source.

Preferably, the oilseed is selected from canola, rape seed, soybeans, sunflower seed, flax seed, mustard seed, cotton seed, hempseed and mixtures thereof, and the animal protein source is selected from whole fish, fish processing waste, fish offal, fish by-catch, squid offal, whole birds without feathers, poultry offal, beef offal, lamb offal and mixtures thereof.

It is desirable the cooked product comprises condensed solubles from the cookingwater-solublized material, the solubles having an enriched soluble nitrogen content, water soluble carbohydrate content, water-soluble antinutritional component, and mineral content.

Moreover, it is preferable the cooked product comprises protein-rich insoJubles -6derived from a cooked admixture of the oilseed and the animal protein source.

It is further preferable the animal feed grade oil has as its protein source fishderived material and comprises: from about 60% to about 92% of total fatty acids as unsaturated fatty acids; from about 8% to about 50% of total fatty acids as fatty acids; from about 3% to about 25% of total fatty acids as n-3 highly unsaturated fatty acids; and, a peroxide value less than about 8 milliequivalents per kg of oil at the time of production.

Desirably, any of the above embodiments includes the subsequent steps of: extracting the protein-rich fraction with a solvent for lipid type materials; and, removing the solvent to obtain a protein concentrate.

In another embodiment of the present invention there is provided a protein concentrate as produced by the processes.

Desirably, the oilseed is greater than 55% dehulled.

It should be noted the animal feed grade oil is an oil derived from raw oilseed; likewise, the edible organic oils may be derived from raw oilseed.

In another development of the present invention, one embodiment provides for a protein and lipid-rich oilseed meal suitable for use in fish and non-human animal diets comprising a heat-treated dehulled oilseed, the oilseed meal having: from about 26% to about 40% protein on a dry weight basis; from about 48% to about 64% protein on a lipid-free dry weight basis; from about 2.4% to about 4.6% methionine and cystine calculated as a percent of said protein; -7from about 3.6% to about 6.1% lysine calculated as a percent of said protein; from about 21% to 52% lipid on a dry weight basis; from about 2% to about 12% crude fibre on a lipid-free dry weight basis.

It will be understood that reference to the above described processes which are suitable for animal and fish feeds, refers to processes which can be used by numerous types of species. For example, depending on the geographic location, the process can be used in fish farming operations for salmon, trout, tilapia, carp, catfish, sea bream and many other warm water as well as cold water species of commercial importance. In the case of animals, the process can be used with conventional farming practices utilizing feeds for poultry, hogs, swine and cattle.

In further explanation of the products and process aspects of the present invention, the solvent used for extracting the mixture obtained from the blended mixture of oilseed and animal offal includes hexane or other compatible solvents used in the animal feed or human food industry.

The ash content in the protein concentrates can be regulated as desired by controlling the concentration of bone in the animal protein source or animal offal.

Thus, the ash can be controlled by using a deboning step to obtain offal with the desired bone content. Bones in wet or dry form of different types of offal can be utilized, with varying degrees of bone coarseness. By way of example, the ash content can thus be controlled by controlling the amount of bone added to the mixture of oilseed and animal offal or animal protein source.

In both the process and product aspects of the present invention, when referring to animal offal such as birds or chickens, it is to be understood that the offal is without bird feathers.

In both the process and product aspects of the present invention, when using dehulled seeds, the term "dehulled" is intended to mean seeds which have substantially all of their hulls removed. However, in many cases, partially dehulled seeds can be employed as otherwise noted- herein, and to this end, dehulled seeds are those which have had at least 55% of their hulls removed.

The above described process can produce various products described hereinafter in greater detail.

As used in the specification, the term "unhydrolyzed" in describing the animal protein source or animal offal refers to animal offal which has less than about by weight of hydrolyzed content, desirably less than 5% and most desirably no hydrolysis whatsoever (fresh, unspoiled). The amount of hydrolyzed content is as close as possible to 0% in order to best achieve the highest nutritive value in the products that are formed.

In this invention, the animal offal is preferably in a particulate form such as that which would be obtained by processing procedures resulting in minced offal. Well known techniques in the offal processing art can be employed to obtain such minced offal.

Brief Description of the Drawing Having generally described the invention, reference will be made to the accompanied drawing which illustrates the preferred embodiments only. Figure 1 is a schematic representation of the process according to the present invention.

Detailed Description of the Invention The steps involved in the process of the invention are broadly represented in Figure 1. In this Figure, there is illustrated a schematic representation of the coprocessing of animal offal(s) with oilseed(s) to yield cold pressed oil indicated as product 1; hulls from dehulled oilseed meats indicated as product 2; nutritionally upgraded oilseed meal produced from heat treated, dehulled and cold pressed oilseed indicated as product 3; animal-feed grade oil indicated as product 4; condensed solubles indicated as product 5; and high nutritive value protein concentrate indicated as product 6. Other products of the invention are obtained by further processing the above-mentioned products as will be described in greater detail hereinafter.

In accordance with certain embodiments of the invention, undehulled oilseed is used in the process. Other embodiments involve dehulled seed and raw seed.

Dehulled seed is preferred when it is desired to feed monogastric species such as -9fish and poultry, and the preferred raw seed used in this embodiment includes canola, sunflower, or delinted cottonseed.

The following examples are presented to describe embodiments of the invention and are not meant to limit the invention unless otherwise stated.

Examples 1 to 10 outlined below described each step involved in the process of the invention: EXAMPLE 1: Animal offal A common batch of whole Pacific herring was used as the main source of animal offal for the project. Soon after the herring were caught, they were rapidly block frozen by McMillan J.S. Fisheries Ltd., Vancouver, BC and stored at -40_C for about 9 months. At this time, about 500 kg of herring were transported to the Department of Fisheries and Oceans, West Vancouver Laboratory where they were held at -20_C until small batches of about 50 kg were partially thawed for each test run. The thawed herring were cold extruded using a Butcher Boy equipped with an auger, cutter knife, and perforated plate having holes with diameter 9.52 mm.

Fresh poultry offal (heads and viscera minus feet) was also used for some trials that involved co-processing the offal with partially dehulled animal feed grade sunflower seed (designated as batch 2 hereinafter). The offal was obtained from West Coast Reduction Ltd., Vancouver, BC and was stored for one night at under cover before being handled as described above for the herring.

EXAMPLE 2: Oilseeds The four oilseeds that have been tested successfully in this project include Goliath canola seed (Cloutier Agra Seeds Inc., Winnipeg, MB), soybeans (InfraReady Products Ltd., Saskatoon SK), sunflower (completely dehulled confectionary grade seed obtained from North West Grain, St. Hilaire, MN, USA (batchl 1) and undehulled animal feed grade seed obtained from Cargill Incorporated, Wayzata, MN, USA; batch and devitalized hemp seed (Seedtec/Terramax, Qu'Appelle, SK sterilized by InfraReady Products Ltd., Saskatoon SK). Delinted glandless cottonseed (California Planting Cottonseed Distributor, Bakersfield, CA,USA) and brown flax (InfraReady Products Ltd., Saskatoon, SK) were also tested in the process. The analytical results pertaining to products based on the former are pending. It was concluded that flax seed would be suitable for the process provided that the seed is almost totally dehulled or the outer mucilage layer of the seed coat is removed through an economical process.

EXAMPLE 3: Heat treatment or micronization of oilseeds In a preferred embodiment of the invention, specially for canola, soya, flax and hemp, an initial heat treatment was performed. The process involved subjecting the whole seeds to infrared energy so that the seed temperature reached 110- 1150C for 90 seconds. Subsequently, the micronized seeds were held for 20-30 min, depending upon the seed source, in an insulated tank where temperatures ranged from 100-1100C (residual cooking conditions). These conditions inactivated enzymes such as myrosinase in canola and trypsin inhibitors in soya as well as peroxidase and cyanogenic glucosides. Further, they ensured devitalization of viable germ tissue in hemp, improved starch digestibility, and destroyed or reduced the concentrations of heat labile antinutritional factors other than those mentioned above.

Sunflower seeds (batches 1 and 2) were not micronized before co-processing with animal offal but the batch 1 seeds were dried to 10% moisture to ensure proper seed storage and facilitate dehulling. Thus, only non-micronized dehulled sunflower seeds were tested in this study.

EXAMPLE 4: Oilseed dehulling Micronized canola, soya, hemp and flax and non-micronized animal feed-grade sunflower were dehulled. The process involved seed sizing, impact dehulling (Forsberg model 15-D impact huller), screening and air classification (Forsberg model 4800-18 screener and screen-aire).

EXAMPLE 5: Oilseed cold-pressing In a preferred embodiment of the invention, the oilseeds (micronized or raw), except soya and micronized dehulled hemp were cold-pressed at a temperature not exceeding 85_C, using a Canadian designed and manufactured laboratory -11 scale Gusta cold press (1 HP Model 11, Gusta Cold Press, St. Andrews, Manitoba, Canada). This served to remove some (dehulled seeds) or a significant proportion (undehulled seeds) of the residual oil (organic human food grade oil) and concomitantly reduced the particle size of the oilseed before it was co-processed with minced animal offal in various proportions (improved the efficiency of the subsequent aqueous extraction of the water soluble antinutritional factors and oligosaccharides present in the oilseed).

In a more preferred embodiment, specially for soya, the particle size was further reduced, using a modified crumbler (model 706S, W.W. Grinder Corp., Wichita, Kansas). This machine was equipped after modification with dual motorized corrugated rolls. One of these had a fixed speed whereas the speed of the other could be varied. For the purpose of this investigation, the variable speed roller was adjusted to rotate much faster than the fixed speed roller to achieve a shearing action.

EXAMPLE 6: Mixing or co-processing step Thawed, ground, whole animal offal (mostly herring, but in two cases poultry offal minus feet, was used) and oilseeds that had been micronized or dried as described in Example 3 or in raw form and either cold pressed or ground as described in Example 5 were first combined in various proportions. In preferred embodiments, the usual percentages of offal to oilseed were 75:25; 50:50; or 25:75 Thereafter, 100 mg of santoquin (antioxidant) per kg of mixture in a marine oil carrier (lg/kg) were added. Then hot water was added to the mixture in such a way that the ratio of water to oil-free dry matter present in the oilseed was maintained between 3-6:1 depending upon the source and proportion of oilseed in the mixture. Both the endogenous water originating from the offal and the exogenous water were considered when calculating the aforementioned ratios.

EXAMPLE 7: Cooking step The mixture obtained from co-processing of animal offal and oilseed (Example 6) was cooked for about 27 min at 90-93°C in the steam jacketed cooker section of a pilot-scale fish meal machine (Chemical Research Organization, Esbjerg, Denmark), that was equipped with a heated auger (it is noteworthy that the -12cooking step could have also been performed by using a heat exchanger with a positive displacement pump or through direct steam injection coupled with processor). The cooking step was undertaken to: minimize the loss of soluble protein through protein denaturation, destroy or reduce the concentration of heat labile antinutritional factors present in the oilseed (especially important when processing non-micronized seeds and micronized soya), liberate the bound cellular water and lipid in the offal and the oilseed, and subject the oilseed to aqueous washing to facilitate removal of the water soluble antinutritional factors originating from this source.

EXAMPLE 8: Pressing step Significant but not total removal of the latter as well as lipid (animal-feed grade product) was accomplished by passing the cooked mixture through the fish meal machine screw press that was equipped with perforated screens and then a laboratory-scale press (Vincent model CP-4; Vincent Corp., Tampa Florida).

Constituents in the water fraction of the press liquids consisted of water soluble carbohydrates such as monosaccharides, disaccharides, or problem sugars like raffinose and stachyose, phenolic compounds, glucosinolates (when canola used), chlorogenic acid (when sunflower used), isoflavones and saponins (when soybeans used) as well as some soluble nitrogen and water soluble vitamins. In preferred embodiments, the presscake in each case was dried in the steam jacketed drier portion of the above-mentioned fish meal machine at 75-83°C to produce dried protein and lipid-rich products.

EXAMPLE 9: Drying step In one preferred embodiment, further drying of the protein products was necessary to reduce their moisture content. The drying was performed for about 30 min to reduce their moisture content to less than 10%. This was accomplished using a custom designed vertical stack (stainless steel mesh trays) pellet cooler that was equipped with two electric base heaters and a top mounted variable speed fan.

The temperature of the upward drawn air was maintained between 70 0 C and 80 0

C

during the process. All protein and lipid sources stemming from the above process, including the cold-pressed oils were further stabilized with santoquin -13- (ethoxyquin). In a more preferred embodiment, specially in the case of the dried protein products, 100 mg of santoquin were added per kg of product in a marine oil carrier (1 g/kg). Then, each of the products was vacuum packaged in oxygen impermeable bags and stored at -20°C pending chemical analysis or their evaluation in a digestibility trial (see below). In another embodiment, specially in relation to the oils, 500 mg of santoquin were added per kg and then each lipid source was stored at 4-5C in 1 L black plastic bottles.

EXAMPLE 10: Separation step In preferred embodiments, the press liquid was separated into water and lipid fractions using an Alpha de Laval batch dairy centrifuge (Centrifuges Unlimited Inc., Calgary, Alberta). Then, the water fraction was condensed to about one third of its original volume using a steam jacketed bowl cooker.

EXAMPLE 11: Preparation of protein concentrates Protein concentrates that are mostly based on protein from canola, soya, sunflower and hemp were prepared by hexane extracting the products that originated from the co-processing of 1:1 combinations of whole herring and each of the preceding oilseeds. In this regard, 200 g of each of the four protein products were extracted four times with hexane (5:1 During each extraction, the mixture was held for 30 min (stirred once after 15 min) before being filtered through Whatman No.1 filter paper in a Buchner funnel. Following hexane extraction, each protein product was placed on a tray that was lined with aluminum foil and then it was air-dried ovemrnight. Then, each product was placed in the pellet cooler described in Example 9, where it was dried at about 70-80 0 C for min to remove any residual traces of hexane.

EXAMPLE 12: In vivo protein digestibility experiments In a preferred embodiment, the in vivo availability (digestibility) of protein in some of the test protein sources that were prepared by co-processing various proportions of whole herring with canola, soya, sunflower and hemp was determined using Atlantic salmon in sea water as the test animal. Two experiments were conducted and the experimental conditions for each are -14provided in the table 1 below, wherein the flow rate of the oxygenated, filtered, ambient sea water was 6 8 L/min, feeding frequency was twice daily, ration was maximum (fish fed to satiation), and the photoperiod was natural.

Table 1.

Variable Experiment 1 Experiment 2 Fish source NorAm Aquaculture, NorAm Aquaculture, Campbell River, BC Campbell River, BC Range in initial mean 76.6-85.8 54.2-61.6 weight (g) Number of fish per tank 15 Tanks per diet 3 3 Stockino density (ko/m 3) <8.6 <6.2 Water temperature 8.9-9.1 9.0-9.5 Salinity 29-31 28-30 Dissolved oxygen (mg/L) 8.5-9.4 7.5-9.0 Fecal collection period 14 13 (days) The design of the digestibility tanks and the fecal collection procedures have been described by Hajen et al. (1993a,b. Aquaculture 112: 321-348). The experimental diets consisted of 29.85% test protein product, 69.65% reference diet, and chromic oxide as the indigestible marker. Table 2 outlined below provides the ingredient and proximate composition of the reference diet used in the digestibility experiments.

Table 2.

Ingredients (g/kg; air-dry basis) LT Anchovy meal 643.2 Blood flour; spray-dried 41.0 Pregelatinized wheat starch 80.9 Raw wheat starch 26.9 Vitamin supplement 1 18.9 Mineral supplement 2 18.9 Menhaden oil; stabilized 3 122.4 Soybean lecithin 9.46 Choline chloride 4.73 Vitamin C, monophosphate 3.38 Permapell 9.46 Finnstim T 14.2 DL-methionine 1.51 Chromic oxide 5.00 Level of: Dry matter 924-926 Protein 452-453 Lipid 184 Ash 118-123 1 The vitamin supplement provided the following amounts/kg of diet on an air-dry basis: vitamin A acetate, 4731 IU; cholecalciferol (D 3 2271 IU; DL-a- tocopheryl acetate 284 IU; menadione, 17.0 mg; D-calcium pantothenate, 159.3 mg; pyridoxine HCI, 46.6 mg; riboflavin, 56.8 mg; niacin, 283.8 mg; folic acid, 14.2 mg; thiamine mononitrate, 53.0 mg; biotin, 1.42 mg; cyanocobalamin (B12), 0.085 mg; inositol, 378.5 mg.

2/ The mineral supplement provided the following (mg/kg diet on an air-dry basis): manganese (as MnSO 4

H

2 71.0; zinc (as ZnSO 4 7H 2 85.2; cobalt (as CoCI2 _6H 2 2.84; copper (as CuS04_5H 2 6.62; iron (as FeS4 _7H 2 94.6; iodine (as K10 3 and KI,1:1), 9.46; fluorine (as NaF), 4.73; selenium (as Na 2 SeO3), 0.19; sodium (as NaCI), 1419; magnesium (as MgSO 4 _7H 2 378; potassium (as K 2 SO4 and K 2 C0 3 1419.

3 Stabilized with 0.5 g santoquin/kg oil.

-16- After adjustment of all experimental diet mashes to a moisture content of they were cold pelleted using a California model CL type 2 pellet mill. Diet particle size was adjusted to suit fish size. The reference and experimental diets that were used in the study were stored at 50C in air-tight containers until required.

The reference and experimental diets (mixture of reference and test diet) and lyophilized fecal samples were analyzed for levels of moisture, protein and chromic oxide at the DFO, West Vancouver Laboratory (WVL) using the procedures described below. Subsequently, the digestibility coefficients for protein were determined for each diet according to Cho et al. (1985. Finfish nutrition in Asia: methodological approaches to research and development. IDRC Ottawa, Ont., 154p.). Then, the digestibility coefficients for each of the protein products themselves were calculated according to Forster (1999. Aquaculture Nutrition 5: 143-145).

The results of chemical analyses of the protein sources used in this study and of the products derived from the co-processing of animal offals (herring or poultry offal) with canola, sunflower, soya and hemp treated as described above are presented in Tables 3-20. The results have been expressed on a dry weight basis and a lipid-free dry weight basis since the mechanical pressing of lipid from the cooked blends of offal and oilseed was variable and not complete. This is a function of the design of the presses and other conventional presses available in industry can be of higher efficiency.

Examples 13 to 16 outlined hereinafter give the results of chemical analyses performed on products obtained in accordance with the process of the invention from: canola and canola-based products, sunflower and sunflower-based products, soya and soya-based products, as well as hemp and hemp-based products. The chemical analyses were performed according to the following methods: Concentrations of protein, moisture, and ash in the protein sources and products that were prepared as well as in all test diets and fecal samples were determined at the Department of Fisheries and Oceans, West Vancouver Laboratory (DFO-WVL) using the procedures described by Higgs et al. (1979. In J.E. Halver, and K. Tiews, eds.

Finfish Nutrition and Fishfeed Technology, Vol. 2. Heenemann Verlagsgesellschaft MbH., Berlin, pp. 191-218).

-17- Similarly, the fatty acid compositions of the cold pressed oils and animal feed grade oils stemming from the press liquids were determined at the same laboratory using the procedures of Silver et al (1993. In S.J. Kaushik and P. Luquet, eds. Fish nutrition in practice. IV h International Symposium on Fish Nutrition and Feeding, INRA, Paris, pp. 459-468).

Moreover, the chromic acid concentrations in diets and lyophilized fecal samples were determined at the DFO-WVL using the methods of Fenton and Fenton (1979. Can. J.

Anim. Sci., 59: 631-634).

Concentrations of crude fibre (AOCS Official Method Ba 6-84), lipid (Troeng, S. 1955.

J.A.O.C.S. 32: 124-126), chlorogenic acid (capillary electrophoresis method developed by M. Marianchuk at the POS Pilot Plant Corp.) and sinapine (capillary electrophoresis method developed by P. Kolodziejczyk et al at the POS Pilot Plant Corp.) in the oilseeds and test protein products as well as measurements of trypsin inhibitor (AOCS Official Method Ba 12-75 reapproved 1997) and urease (AOCS Official Method Ba 9-58 reapproved 1993) activities in soya and sunflower seeds and protein products were determined at the POS Pilot Plant Corp., Saskatoon, SK.

according to the methods cited in the parentheses.

Determinations of the amino acid concentrations in the oilseeds and test protein products were conducted by AAA Laboratory, Mercer Island, WA, USA using the general procedures described by Mwachireya et al. (1999. Aquaculture Nutrition 73-82).

Levels of phytic acid in all oilseeds and in the products derived from the coprocessing of oilseeds and animal offal were determined by Ralston Analytical Laboratories, Saint Louis, MO using the procedures described by Forster etal. (1999.

Aquaculture 179: 109-125).

Mineral concentrations in the oilseeds and the protein products were determined by Norwest Labs, Surrey, BC using plasma spectroscopy (Higgs et al, 1982.

Aquaculture 29: 1-31).

Concentrations of glucosinolate compounds (total of all the different types of glucosinolates) present in canola and canola- based products were measured by Dr.

-18- Phil Raney, of Agriculture Agri-Food Canada, Saskatoon, SK according to the methods of Daun and McGregor (1981. Glucosinolate Analysis of Rapeseed (Canola). Method of the Canadian Grain Commission Revised Edn. Grain Research Laboratory, Canadian Grain Commission, Winnipeg, Manitoba, Canada).

Measurements of soy isoflavones namely, daidzein, glycitein, genistein, and saponins were conducted by Dr. Chung-Ja C. Jackson, of the Guelph Center for Functional Foods, University of Guelph Laboratory Services and have been reported here as the total for the preceding compounds (the methodology in each case is the subject of a patent application and hence has not been published).

EXAMPLE 13: Results obtained for canola and canola-based products Table 3 outlined below gives the percentages of extensively dehulled and partially dehulled Goliath canola seed and of hulls in relation to seed size after dehulling by Forsberg Incorporated, Thief River Falls, MN.

Table 3.

Seed size/fraction Weight (kg) Extensively 35.8 39.4 dehulled; large 1/ Extensively 10.8 11.8 dehulled; small 1/ Partially dehulled; 20.4 22.4 large 2 Partiall dehulled; 14.3 15.7 small Hulls; small 3/ 3.33 3.66 Hulls; large 3 6.49 7.13 Total 91.1 100 1/ The extensively dehulled canola as identified visually by the lack of hulls in the material was used in the tests reported below (referred to as dehulled canola) -19- 2 The partially dehulled canola could be subjected to further dehulling, directed into ruminant diets, and/or mixed at a low proportion with animal offal and then coprocessed to create a nutritionally upgraded protein source for monogastrics.

3 The hulls contained little visible evidence of canola meats and had low density.

Table 4 gives the percentages of presscake and oil obtained after cold pressing raw, undehulled and micronized, dehulled Goliath canola seed using a laboratory scale Gusta press.

Table 4.

Raw, undehulled Micronized, dehulled Fr n canola seed canola seed Fraction Presscake 68.3 84.0 Oil 31.7 16.0 Total 100 100 Table 5 sets out the initial ratios of water from endogenous and exogenous sources to oilseed lipid-free dry matter content and percentage yields (air-dry product, moisturefree product, and lipid-free dry weight product) from the co-processing of different blends of whole herring (WH) with dehulled, micronized (DC) and undehulled raw Goliath canola seed (URC).

Table Protein product/ Initial ratio of hot Air-dry Moisture- Lipid-free water to oilseed product free product dry product lipid-free dry matter (w/w) WH75DC25 5:1 29.4 27.0 19.4 WH50DC50 5:1 32.7 31.1 20.4 WH37.5DC62.5 5:1 34.8 31.8 20.0 WH75URC25 4.5:1 30.5 27.1 19.0 WH50URC50 5:1 30.9 29.8 21.3 WH25URC75 5:1 29.6 28.6 20.5 1/ Numbers following WH, DC, and URC refer to initial percentages of these products in the herring/canola seed blends (canola seed was cold pressed to remove a significant portion of the oil and reduce the particle size of the starting material before blending with herring and santoquin; 0.1g/kg of mixed product before water addition) before their co-processing using cooking temperatures of 90-93_C and drying temperatures of 77-83_C.

In Table 6, the concentrations of proximate constituents including crude fibre (CF) as well as phytic acid total glucosinolates and sinapine in whole herring dehulled micronized cold pressed Goliath canola undehulled raw cold pressed Goliath canola (URC), and six protein products produced by the coprocessing of different proportions of WH with either DC or URC (expressed on a dry weight basis, DWB or lipid-free dry weight basis, LFDWB) are provided. The composition of a seventh protein product that was produced by hexane extraction of WH50DC50 is also shown (WH50DC50-hexane) together with the apparent protein digestibility coefficients for some of the products (Atlantic salmon in sea water used as the test animal) is also provided.

-21 Table 6.

Parameter WH DC URC WH75 WH50 WH50 WH37.5 WH75 WH50 DC50 DC50 DC62.5 URC25 URC5 (hexane) 0 Dry matter (g/kg) 286 954 936 918 952 928 914 890 966 968 Protein (g/kg) -DWB 488 279 348 529 456 693 416 525 414 404 -LFDWB 870 515 469 735 696 724 662 748 578 564 Lipid (g/kg) -DWB 439 458 258 280 345 42.5 372 298 284 284 Ash (g/kg) -DWB 70.3 48 60.9 81 67.1 97.2 63 77.1 78.8 73.5 -LFDWB 125 88.6 82.1 113 102 102 100 110 110 103 CF (g/kg) -DWB 28.3 66.5 21 24.7 38.3 28.9 69.2 76.4 -LFDWB 52.2 89.6 29.2 37.7 40 46 96.6 107 PA (g/kg) -DWB 28.2 33.9 15.6 22.9 25.5 14.2 26 30.7 -LFDWB 52 45.6 21.6 35 40.6 20.2 36.3 42.9 TG (gmoles/g) -DWB 10.8 17.8 1.09 1.26 0.92 0.44 0.9 1.06 -LFDWB 19.9 24 1.52 1.92 1.47 0.63 1.26 1.49 Sinapine(g/kg) -DWB 11.2 13.1 3.16 4.94 5.8 2.92 5.18 5.68 -LFDWB 20.7 17.7 4.39 7.54 9.23 4.16 7.23 7.94 In vivo protein 88.9 94.4 94.9 94.4 96.4 digestibility 1 Not determined c -22- Table 7 provides the concentrations of essential amino acids of protein) and selected minerals (pg/g of lipid-free dry matter) in whole herring micronized, dehulled, cold pressed Goliath canola undehulled, raw cold pressed Goliath canola (URC), and six protein products produced by the co-processing of different propotions of WH with either DC or URC. The amino acid and mineral concentrations in a seventh protein product, produced by hexane extraction of WH50DC50 are also shown (WH50DC50-hexane).

-23- Table 7.

Parameter WH DC URC WH75 WH50 WH50 WH37.5 WH75 WH50 DC50 DC50 DC62.5 URC2 URC5 (hexane) 5 0 A) Essential amino acids Arginine 6.66 7.09 7.23 7.44 7.5 6.93 Histidine 1.97 2.84 2.62 2.69 2.69 2.59 Isoleucine 4.56 4.28 4.81 4.78 4.71 4.51 Leucine 8.4 7.47 8.15 8.22 8.01 7.71 Lysine 5.47 3.87 4.92 4.85 7.01 4.4- Methionine 3.97 4.55 4.54 4.63 4.25 4.47 Cystine Phenylalanine 7.55 7.26 8.08 8.14 7.93 7.54 Tyrosine Threonine 4.97 4.62 4.83 4.89 4.73 4.61 Tryptophan 1.51 1.72 1.69 1.63 0.92 1.69 Valine 5.51 5.34 5.66 5.75 5.23 5.36 B) Minerals Calcium 30303 4061 5183 23905 14594 16202 12195 22088 14458 10244 Phosphorus 19073 18760 17278 23299 21971 23746 20384 21127 20675 20777 Magnesium 1961 7929 7631 4388 5934 6921 6098 4161 6289 8599 Sodium 5704 <100 <100 3026 1443 1598 1220 2081 772 495 Potassium 14260 18566 18142 12104 12348 14293 12544 10244 11234 12019 Copper 5.2 3.09 <1.00 15.4 6.09 12.0 8.36 11.4 10.7 9.81 Zinc 101 70.8 66.7 116 101 106 79.6 96.2 74.6 71.1 -24- Table 8 sets out the percentages of selected fatty acids and of saturated, unsaturated, and n-3 highly unsaturated fatty acids (n-3 HUFA; 20:5 (n- 3) 22:6 in whole herring undehulled raw cold pressed Goliath canola (URC), and the press lipids resulting from th e co-processing of different proportions of WH with DC or URC.

Table 8.

Lipid source WH URC WH75 WH50 WH37.5 WH75 WH50 Fatty acid DC25 DC50 DC62.5 URC25 URC50 18:1(n-9) 18.9 62.7 35.7 49.9 55.0 22 25.7 18:2(n-6) 0.74 21.4 7.34 15.0 17.5 10.3 17 18:3(n-3) 0.12 8.79 3.28 0.34 7.40 2.67 4.28 20:5(n-3) 9.66 0,00 15.4 3.75 2.71 4.21 5.83 22:6(n-3) 8.96 0.00 7.00 4.11 1.38 6.69 0 Total Saturated 22.2 4.32 20,3 12.4 9.88 22.1 20.3 Total Unsaturated 77.8 95.7 79.7 87.6 90.1 77.9 79.7 Total 4.82 21.7 8.56 15.5 17.9 14.1 Total 31.3 9.96 28.0 14.7 11.8 21.7 15.3 Total n-3 HUFA 18.6 0 22.4 7.86 4.09 10.09 5.813 -26- EXAMPLE 14: Results obtained for sunflower and sunflower-based products In Table 9, initial ratios of water from endogenous and exogenous sources to oilseed lipid-free dry matter and percentage yields (air-dry product, moisture-free product, and lipid-free dry weight product) from the co-processing of different blends of whole herring (WH) or poultry offal (PO) with dehulled, raw sunflower seed, batch 1 (DRSFi) or batch 2 (DRSF 2 are provided.

Table 9.

Protein product Initial ratio of hot Air-dry Moisture- Lipid-free water to oilseed product free dry product lipid-free dry matter product 1 25 5:1 30.4 28.2 19.7 1 50 3:1 31.6 29.0 19.4 WH25DRSF175 3:1 31.7 31.1. 19.9 2 50 6:1 46.9 43.0 31.3 1 Numbers following WH, DRSF and PO refer to initial percentages of these products in the herring/sunflower seed and poultry/sunflower seed blends (sunflower seed was cold pressed to remove a significant portion of the oil and reduce the particle size of the starting material before blending with herring or poultry and santoquin; 0.1 g/kg of mixed product before water addition) before their co-processing using cooking temperatures of 90-93_C and drying temperatures of 77-83_C.

Table 10 gives the concentrations of proximate constituents including crude fibre phytic acid trypsin inhibitor activity urease activity (UA) and chlorogenic acid (CA) content in whole herring poultry offal dehulled, raw cold pressed sunflower, batch 1 (DRSF1), and five protein products produced by the co-processing of different proportions of WH or PO with either DRSF, or dehulled, raw cold pressed sunflower, batch 2 (DRSF 2 (expressed on a dry weight basis, DWB or lipid-free dry weight basis, LFDWB). The composition of a sixth protein product that was produced by hexane extraction of WH50DRSF 1 50 is also shown (WH50DRSFi50-hexane) together with the apparent protein digestibility coefficients for some of the products (Atlantic salmon in sea water used as the test animal).

-27- Table Parameter WH PO DRSF 1 WH75 WH50 WH50 WH25

DRSF

1 25 DRSF 1 50 DRSFi50 DRSF 1 75 DRSF 2 (hexane) Dry matter(g/kg 286 328 938 928 919 930 981 918 Protein (g/kg) 488 370 351 535 479 695 441 382

-DWB

-LFDWB 870 673 594 766 715 718 689 525 Lipid (g/kg) -DWB 439 451 409 302 330 32.2 360 271 Ash (g/kg )-DWB 70.3 104 50.7 126 118 95.8 115 58.9 -LFDWB 125 189 85.8 181 176 99 180 80.8 CF (p/kg) -DWB 34 19.2 32.4 37.5 27.2 124 -LFDW B 57.5 27.5 48.4 38.8 42.5 170 PA (g/kg) -DWB 31.4 14.2 23.3 30.7 25.9 -LFDWB 53.2 20.4 34.8 47.9 35.6 TI (TIA units/g) 1603 1766 1730 1268

LFDWB

UA (ApH) 0.06 0.03 0.02 0.01 CA(g/kg) -DWB 14.9 2.6 5.65 8.58 6.22 -LFDWB 27.7 3.72 8.43 13.4 8.53 In vivo protein digestibility 97.6 97.1 Not determined 21 DRSF2 co-processed with PO was pressed, partially dehulled animal feed grade with a DM, protein, lipid, ash and crude fibre content (g/kg expressed on a dry weight basis except DM) of 918, 379, 211, 59.4, and 123, respectively.

-28- Table 11 gives the concentrations of essential amino acids of protein) and selected minerals (pg/g of lipid-free dry matter) in whole herring poultry offal dehulled, raw, cold pressed sunflower, batch 1 (DRSFi), and four protein products produced by the co-processing of different proportions of WH or PO with either DRSF 1 or DRSF 2 The concentrations in a fifth protein product, produced by hexane extraction of WH50DRSF 1 50, is also shown (WH50DRSF,50-hexane).

-29- Table 11 Parameter WH PO DRSF1 WH75 WH50 WH50 WH25 P050

DRSF

1 25 DRSF,50 DRSF,50 DRSF,75 DRSF 2 (hexane) A) Essential amino acids Arginine 6.66 8.11 10.6 7.66 8.58 8.64 9.16 8.52 Histidine 1.97 1.91 2.59 2.34 2.42 2.40 2.41 2.56 Isoleucine 4.56 3.19 4.45 4.28 4.45 4.52 4.38 4.56 Leucine 8.40 5.88 6.32 7.16 6.96 7.11 6.57 6.95 Lysine 5.47 5.28 3.67 6.88 5.57 5.57 4.30 4.59 Methionine Cystine 3.97 3.16 3.61 3.71 3.41 3.61 3.42 3.25 Phenylalanine Tyrosine 7.55 5.45 7.66 7.46 7.65 7.82 7.55 7.72 Threonine 4.97 3.67 4.15 4.40 4.17 4.10 4.06 3.99 Tryptophan 1.51 0.75 1.28 1.27 0.78 1.27 1.03 1.40 Valine 5.51 4.03 5.19 5.09 5.29 4.86 5.08 4.81 B) Minerals Calcium 30303 1 1930 33810 15055 14999 10226 12420 Phosphorus 19073 22188 29950 25011 23221 23573 15843 Magnesium 1961 10805 4493 7503 7544 9987 4992 Sodium 5704 19.8 2223 1454 1378 836 852 Potassium 14260 23090 11085 14406 15110 15036 9894 Copper 5.20 39.1 21.6 36.5 28.9 37.0 39.9 Zinc 101 124 99.0 118 124 123 93.0 "Not determined.

2 Values for essential amino acids were derived from unpressed DRSFi.

3 DRSF 2 co-processed with PO was partially dehulled animal feed grade with a DM, protein, lipid, ash and crude fibre composition (g/kg expressed on a dry weight basis except DM) of 918, 379, 211, 59.4, and 123, respectively.

In Table 12, percentages of selected fatty acids and of saturated, unsaturated, and n-3 highly unsaturated fatty acids (n-3 HUFA; 20:5 22:6 in whole herring poultry offal dehulled, raw, cold pressed sunflower, batch 1 (DRSFI), and the press lipids resulting from the co-processing of different proportions of WH or PO with DRSF 1 or dehulled, raw, cold pressed sunflower, batch 2 (DRSF 2 -31 Table 12 Lipid source Fatty acid WH PO DRSF 1 WH75 WH50 WH25 P050

DRSF

1 25 DRSF 1 50 DRSF 1 75 DRSF 2 502 18:1(n-9) 18.9 39.9 9.39 21.6 18.2 17.9 18:2(n-6) 0,74 17.6 76.6 22.4 25.3 38.7 18:3(n-3) 0.12 2.56 0.11 4.28 0.42 0.80 20:5(n-3) 9.66 0.00 0.00 3.15 3.67 0.00 22:6(n-3) 8.96 0.00 0.00 6,04 7.87 0.00 Total Saturated 22.2 33.9 12.1 16.9 14.5 37.7 Total Unsaturated 77.8 66.17 87.9 83.1 85.5 62.3 Total 4.82 17.8 76.6 29.5 36.3 38.7 Total 31.3 2.56 0.12 19.2 17.7 0.80 Total n-3 HUFA 18.6 0.00 0.00 9.19 11.5 0.00 1 Not determined.

2/ DRSF 2 co-processed with PO was partially dehulled animal feed grade with a DM, crude protein,lipid, ash and crude fibre content (g/kg expressed on a dry weight basis except DM) of 918, 379, 211, 59.4, and 123, respectively.

-32- EXAMPLE 15: Results obtained for soya and soya-based products In Table 13, the initial ratios of water from endogenous and exogenous sources to oilseed, lipid-free dry matter and percentage yields (air-dry product, moisture-free product, and lipid-free dry weight product) from the co-processing of different blends of whole herring (WH) with dehulled, micronized (DSY) and undehulled raw soya seed (URSY).

Table 13.

Protein product1/ Initial ratio of hot Air-dry Moisture- Lipid-free water to oilseed product free product dry product lipid-free dry matter (w/v) WH75DSY25 5:1 14.2 13.6 10.3 WH50DSY50 5:1 36.7 34.9 26.4 WH25DSY75 4:1 48.3 43.8 32.7 WH75URSY25 5:1 20.7 19.1 15.0 WH50URSY50 5:1 29.9 27.4 21.1 WH25URSY75 4:1 43.8 38.4 33.7 1/ Numbers following WH, DSY and URSY refer to initial percentages of these products in the herring/soya blends (soya seed was ground to reduce the particle size of the starting material before blending with herring and santoquin; 0.1g/kg of mixed product before water addition) prior to their co-processing using cooking temperatures of 90-93 _C and drying temperatures of 77-83 _C.

Table 14 shows the concentrations of proximate constituents including crude fibre (CF) as well as phytic acid total saponins, total isoflavones (TIF), urease activity -33and trypsin inhibitor activity (TI) in whole herring dehulled, micronized,, soya (DSY), undehulled, raw soya (URSY), and six protein products produced by the co-processing of different proportions of WH with either DSY or URSY (expressed on a dry weight basis, DWB or lipid-free dry weight basis, LFDWB). The composition of a seventh protein product that was produced by hexane extraction of WH50DSY50 is also shown (WH50DSY50-hexane) together with the apparent protein digestibility coefficients for some of the products (Atlantic salmon in sea water used as the test animal).

34 Table 14 Parameter WH DSY URSY WH75 WH50 WH50 WH25 WH75 WH50 DSY50 DSY50 DSY75 URSY25 URSY5 (hexane) 0 Dry matter (g/kg) 286 921 897 956 950 936 907 921 916 878 Protein (g/kg) 488 396 334 526 531 647 507 497 429 388

DWB

-LFDWB 870 522 434 696 701 668 680 633 557 504 Lipid (g/kg) 439 242 230 244 242 30.1 254 215 230 232

DWB

Ash (g/kg) 70.3 50.3 57.1 77.2 59.4 71.2 52.2 85.8 66,4 56.8

DWB

-LFDWB 125 66.4 74.2 102 78.4 73.4 70.0 109 86.2 74.0 CF (g/kg) 16.2 44.6 16.0 16.3 18.7 19.3 46.6 67.5 82.2

DWB

-LFDWB 21.3 57.9 21.2 21.5 19.3 25.9 59.3 87.6 107 PA (g/kg) 14.9 20.0 9.87 11.9 12.9 12.5 15.9 17.2

DWB

-LFDWB 19.6 25.9 13.1 15.7 17.3 15.9 20.7 22.4 Saponins (mg/g) 1.60 0.71 1.02 1.18

-DWB

-LFDWB 2.11 0.94 1.35 1.58 TIF(_g/g) 2305 899 1402 1622

DWB

-LFDWB 3041 1189 1850 2174 UA 0.01 2.48 0.02 0.01 0.02 0.09 0.28 0.35 TI (TIA units/g) 7813 101563 871 1017 553 1902 8296 11138

LFDWB

In vivo protein 96.2 94.2 93.5 88.2 digestibility 1 Not determined Table 15 provides concentrations of essential amino acids of protein) and selected minerals (pg/g of lipid-free dry matter) in whole herring dehulled, micronized, soya (DSY), and three protein products produced by the coprocessing of different proportions of WH with DSY. The concentrations in a fourth protein product, produced by hexane extraction of WH50DSY50, is also shown (WH50DSY50-hexane).

-36- Table Parameter WH DSY WH75 WH50 WH50 DSY50 DSY50 (hexane) A) Essential amino acids Arginine 6.66 7.57 7.39 7.17 7.64 7.38 Histidine 1.97 2.48 2.45 2.42 2.49 2.47 Isoleucine 4.56 4.65 4.67 4.60 4.83 4.57 Leucine 8.40 7.53 7.66 7.48 8.00 7.58 Lysine 5.47 6.14 7.13 6.70 6.72 6.52 Methionine Cystine 3.97 2.46 3.30 2.70 3.20 2.97 Phenylalanine Tyrosine 7.55 8.56 8.21 8.27 8.78 8.47 Threonine 4.97 4.21 4.57 4.37 4.44 4.30 Tryptophan 1.51 1.45 1.38 1.31 1.20 1.35 Valine 5.51 4.54 5.26 5.04 4.79 4.99 B) Minerals Calcium 30303 2637 22138 14304 9958 8646 Phosphorus 19073 9339 19648 14998 11897 12385 Magnesium 1961 3638 2684 2597 2324 2971 Sodium 5704 <5.00 2228 1290 1157 668 Potassium 14260 27646 17157 16942 13769 17587 Copper 5.20 21.6 36.7 26.7 23.6 27.2 Zinc 101 57.3 75.3 65.5 65.6 67.8 -37- Table 16 provides the percentages of selected fatty acids and of saturated, unsaturated, and n-3 highly unsaturated fatty acids (n-3 HUFA; 20:5 (n-3) 22:6 in whole herring micronized, dehulled, soya (DSY), undehulled, raw soya (URSY), and the press lipids resulting from the co-processing of different proportions of WH with DSY or URSY.

-38- Table 16.

Lipid source Fatty acid WH DSY URSY WH75 WH50 WH25 WH75 WH50 DSY50 DSY75 URSY25 URSY50 18:1(n-9) 18.9 17.8 17.4 17.9 18.86 15.8 18.4 14.5 13.1 18:2(n-6) 0.74 57.5 57.2 6.39 10.1 22,8 8.24 13.0 25.2 18:3(n-3) 0.12 9.79 10,2 2.67 2.19 4.38 2.99 2.79 4.60 20:5(n-3) 9,66 0.00 0.00 11.2 6.92 5.58 10.4 7.85 5.17 22:6(n-3) 8,96 0.00 0.00 8.55 8.11 6.77 9.10 8.33 6.29 Total Saturated 22.2 13.4 13.3 22.1 25.7 22.8 21.0 26.4 26.8 Total Unsaturated 77.8 86.6 86.7 77.9 74.3 77.2 79.0 73.6 73.2 Total 4.82 57.9 57.8 9.26 10.1 22.8 8.24 13.0 25.2 Total 31.3 10.0 10.4 29.3 25,6 24.5 32.7 27.5 22.1 Total n-3 HUFA 18.6 0.00 0.00 19.7 15.0 12.3 19.5 16.2 11.5 -39- Example 16: Results obtained for hemp and hemp-based products.

In Table 17, the initial ratios of water from endogenous and exogenous sources to oilseed lipid-free dry matter and percentage yields (air-dry product, moisture-free product, and lipid-free dry weight product) from the co-processing of different blends of whole herring (WH) with dehulled, sterilized (DHP) and undehulled sterilized hemp seed (UHP).

Table 17.

Protein product Initial ratio of hot Air-dry Moisture- Lipid-free dry water to oilseed lipid- product free product product free dry matter WH75DHP25 5:1 3.04 2.93 2.80 WH50DHP50 4:1 20.4 19.9 15.1 WH25DHP75 3:1 37.3 32.6 23.2 WH75UHP25 5:1 15.0 14.7 11.9 WH50UHP50 5:1 36.9 36.4 31.4 WH25UHP75 4:1 40.3 39.7 34.2 Numbers following WH, DHP and UHP refer to initial percentages of these products in the herring/hemp blends (UHP seed was cold pressed to remove a significant portion of the oil and to reduce the particle size of the starting material before blending with herring and santoquin; 0.1g/kg of mixed product before water addition) prior to their co-processing using cooking temperatures of 90-93_C and drying temperatures of 77-83 _C.

Table 18 gives the concentrations of proximate constituents including crude fibre (CF) as well as phytic acid (PA) in whole herring dehulled, sterilized hemp (DHP), cold pressed undehulled, sterilized hemp (UHP), and six protein products produced by the co-processing of different proportions of WH with either DHP or UHP (expressed on a dry weight basis, DWB or lipid-free dry weight basis, LFDWB). The composition of a seventh protein product that was produced by hexane extraction of WH50DHP50 is also shown (WH50DHP50-hexane) together with the apparent protein digestibility coefficients for some of the products (Atlantic salmon in sea water used as the test animal).

-41 Table 18.

Parameter WH DHP UHP WH75 WH50 WH50 WH25 WH75 WH50 DHP50 DHP50 DHP75 UHP25 UHP50 (hexane) Dry matter (g/kg) 286 963 952 976 975 969 874 983 986 986 Protein (g/kg) 488 313 311 579 575 721 533 504 429 448

-DWB

-LFDWB 870 632 399 765 757 746 750 625 498 520 Lipid (g/kg) -DWB 439 505 221 243 240 33.4 289 193 138 138 Ash (g/kg) -DWB 70.3 59.2 63.6 99.2 88.7 118 87.6 77.4 116 117 -LFDWB 125 120 81.6 131 117 122 123 95.9 135' 136 CF (g/kg) -DWB 44.1 251 14.7 39.1 51.9 52.5 153 237 239 -LFDWB 89.2 322 19.4 51.4 53.7 73.9 189 275 277 PA (g/kg) -DWB 37.5 33.7 12.5 35.2 47.7 15.3 25.3 24.6 -LFDWB 75.7 43.3 16.5 46.3 67.1 18.9 29.3 28.6 In vivo protein digestibility 96.1 -99.9 Not determined -42- Table 19 shows the concentrations of essential amino acids of protein) and selected minerals of lipid-free dry matter) in whole herring dehulled, sterilized hemp (DHP), and three protein products produced by the co-processing of different proportions of WH with DHP. or UHP. The concentrations in a fourth protein product, produced by hexane extraction of WH50DHP50, are also shown (WH50DHP50-hexane).

-43- Table 19 Parameter WH DHP WH75 WH50 WH50 DHP50 DHP50 (hexane) A) Essential amino acids Arginine 6.66 14.0 8.48 10.4 10.6 11.7 Histidine 1.97 2.81 2.53 2.58 2.62 2.71 Isoleucine 4.56 4.24 4.97 4.72 4.79 4.54 Leucine 8.40 6.72 8.32 7.70 7.90 7.31 Lysine 5.47 3.81 7.93 6.45 6.39 5.35 Methionine Cystine 3.97 4.11 4.11 4.08 4.02 3.91 Phenylalanine Tyrosine 7.55 8.41 8.62 8.48 8.68 8.52 Threonine 4.97 3.71 4.80 4.37 4.36 4.06 Tryptophan 1.51 0.40 0.41 0.75 1.39 0.75 Valine 5.51 4.97 5.58 5.37 5.19 5.20 B) Minerals Calcium 30303 1792 35867 16734 17616 7789 Phosphorus 19073 31048 29641 28340 27652 31219 Magnesium 1961 14202 3668 8772 8531 12375 Sodium 5704 37.8 2558 1646 1708 1162 Potassium 14260 18880 10882 11876 13559 14419 Copper 5.20 30.8 18.7 22.5 26.5 25.4 Zinc 101 169 101 125 141 154 -44- Table 20 sets out the percentages of selected fatty acids and of saturated, unsaturated, and n-3 highly unsaturated fatty acids (n-3 HUFA; 20:5 (n- 3) 22:6 in whole herring dehulled, sterilized hemp (DHP), undehulled, sterilized hemp (UHP), and the press lipids resulting from the coprocessing of different proportions of WH with DHP or UHP.

Table Lipid source WH DHP UHP WH75 WH50 WH25 WH75 WH50 Fatty acid DHP25- DHP50 DHP75 UHP25 UHP50 18:1(n-9) 18.9 5.48 7.72 8.30 8.63 6.51 15.5 17.1 14.2 18:2(n-6) 0.74 57.7 56.4 30.9 42.7 49.8 18.7 18.9 33.6 18:3(n-3) 0.12 19.8 19.0 10.9 15.0 16.8 7.12 7.32 11.5 20:5(n-3) 9.66 0.16 0.02 4.30 2.72 1.55 4.63 3.46 0.05 22:6(n-3) 8.96 0.00 0.00 3.94 2.68 1.91 7.05 5.94 3.00 Total Saturated 22.2 12.5 10.0 24.0 16.4 14.6 20.8 21.4 16.7 Total Unsaturated 77.8 87.5 90.0 76.0 83.6 85.4 79.2 78.6 83.3 Total 4.82 57.7 60.6 31.6 42.8 49.9 21.8 20.9 36.5 Total 31.3 20.3 19.1 23.4 24.2 22.1 25.9 22.6 18.3 Total n-3 HUFA 18.6 0.16 0.02 8.25 5.39 3.46 11.7 9.40 3.05 46 The co-processing of animal offal with the foregoing oilseeds pretreated using the methods according to the present invention resulted in nutritionally upgraded protein sources suitable for use.

The yields of these protein sources were good for all canola and sunflower-based products and this was also true for the soya and hemp-based products when higher concentrations 50% in initial mixture) of these treated oilseeds were used. All of the yields were likely underestimated of true values owing to the difficulty in quantitatively collecting all of the material from the drier portion of the fish meal machine.

The oilseed-based protein products contained high concentrations of protein that was highly bioavailable to salmon (generally 89% to 100% of the protein was noted to be digestible in Atlantic salmon held in sea water depending upon the source and percentage of the oilseed in the initial mixture of offal and oilseed and the pretreatment of the latter and the offal before their co-processing). Moreover, these protein products had significantly reduced concentrations of all heat labile and water soluble antinutritional factors except phytic acid relative to their respective initial levels in the oilseeds. Phytic acid was concentrated during the co-processing of offal with oilseed and the extent depended upon its initial concentration in the oilseed used in the process.

The fatty acid compositions of the animal feed grade lipid sources produced by the process largely reflected the fatty acid compositions and lipid levels contributed by the different proportions of the animal offal and oilseed used initially in the process. This provides considerable scope to produce specially designed lipid sources that are tailored to meet the fatty acid needs of various animal species.

The cold-pressing of oilseeds before they are blended with animal offal yielded high quality economically valuable human food grade oils whose fatty acid compositions can be varied, depending upon market requirements and the selection of the oilseed or combination of oilseeds that are used in cold pressing. The high value of the cold pressed oils which can be generated in greater quantities wen undehulled seeds rather than dehulled seeds are cold pressed will contribute to the overall economic viability of the co-processing of animal offals with oilseeds.

47 The hulls resulted from the dehulling of the oilseeds used in this study and the condensed solubles produced by co-processing animal offal(s) with oilseed(s) likely will be excellent organic fertilizer constituents. This is because they collectively contain soluble protein, some lipid and minerals and other components that can be degraded by aerobic or anaerobic bacterial processes into value-added fertilizer products making the overall process described herein economically viable.

The rapid heat treatment of oilseeds to inactivate enzymes like the protease inhibitors in soya and destruct heat labile antinutritional components coupled with the dehulling of oilseeds yield protein and lipid-rich products that potential can be used directly in high energy feeds such as those destined for aquatic species like salmon (salmon grower-diets frequently contain 25-35% lipid on an air-dry basis and they are produced by extrusion processing technology).

The word 'comprising' and forms of the word 'comprising' as used in this description and in the claims does not limit the invention claimed to exclude any variants or additions.

Claims (26)

1. A process for the preparation of human grade oil, nutritionally upgraded oil seed meals, protein concentrates, lipid rich meals having a reduced fibre content and animal feed grade oils for use in fish or other non-human animal diets comprising providing a source of oilseed; pretreating said oilseed by one of: heat treating sufficiently to substantially reduce the concentration of at least some heat sensitive enzymes or antinutritional components normally present in said oilseed to obtain heat-treated seed that is comminuted by cold pressing to obtain human-grade oil or grinding; or, (ii) cold pressing said oilseed under conditions to substantially reduce particle size of said oilseed and obtain pressed raw seeds; and, blending said pretreated oilseed with unhydrolyzed comminuted or chopped animal protein sources together with an antioxidant, and when required to facilitate cooking and the extensive removal of water soluble antinutritional factors and carbohydrates in said oilseed, including water to produce a mixture thereof; cooking said mixture under conditions to substantially improve protein digestibility, and substantially free cellular water present in said animal protein source and facilitate separation of protein from the lipid in said animal protein source and said oilseed to obtain a cooked mixture; and, separating said cooked mixture into a stickwater fraction, a mostly insoluble protein-rich fraction, and an animal feed grade oil fraction.

2. A process as defined in claim 1, wherein said pretreating comprises a drying step, by cold pressing or by heat treating, and the dried heat treated seed is dehulled to produce a meat fraction and a hull fraction with further cold pressing being carried out on the meat fraction sufficiently to yield a

20-12-05; 8:27AM;MSJ Mel 2 9296 3999 2/ 2 o -49- C" high value human grade oil which is separated and a protein-and-lipid-rich Smeal, and blending this resulting meal with said animal protein source before cooking. 0 3. A process as defined in claim 1 or 2, wherein the pretreating and blending steps include: drying said seed to reduce its moisture content to below about 10% by 00 weight to obtain a dried seed or subjecting said oilseed to heat treatment C] under conditions selected to substantially deactivate, destroy or reduce the o concentration of at least some of the antinutritional components normally C- 10 present in oilseed to produce a heat-treated seed; blending said oilseed and said animal protein source in a weight ratio of about 10:90 to about 90:10, to form a mixture thereof. 4. A blended cooked feed product comprising: an oilseed-derived component selected from at least one of cooking water solubilized material, protein-rich insolubles and feed grade oil; wherein said cooking water solubilized material, protein-rich insolubles and feed grade oil are produced according to the process of claim 1; and, an unhydrolyzed animal protein-source-derived component selected from at least one of cooking-water-solublized material, protein-rich insolubles and feed grade oil wherein said cooking water solubilized material, protein-rich insolubles and feed grade oil are produced according to the process of claim 1. The feed product of claim 4, wherein said cooked product comprises an animal feed grade oil comprising a cooked admixture of lipids from at least one of an oilseed and an animal protein source. 6. The feed product of claim 4, wherein said cooked product comprises condensed solubles from said cooking-water-solublized material, said solubles having an enriched soluble nitrogen content, water soluble COMS ID No: SBMI-02238248 Received by IP Australia: Time 08:30 Date 2005-12-20 o carbohydrate content, water-soluble antinutritional component, and mineral content. 0 7. The feed product of claim 4, wherein said cooked product comprises Sprotein-rich insolubles derived from a cooked admixture of said oilseed and said animal protein source. 8. The feed product of claim 5, wherein the animal feed grade oil has as its (0 protein source fish-derived material and comprises: 0 from about 60% to about 92% of total fatty acids as unsaturated fatty Sacids; from about 8% to about 50% of total fatty acids as fatty acids; from about 3% to about 25% of total fatty acids as n-3 highly unsaturated fatty acids; and, a peroxide value less than about 8 milliequivalents per kg of oil at the time of production. 9. The process of claim 1, 2 or 3 including the subsequent steps of: extracting said protein-rich fraction with a solvent for lipid type materials; and, removing said solvent to obtain a protein concentrate. The protein concentrate as produced by the process of claim 9. 11. The blended cooked feed product of claim 4, wherein said product comprises a protein concentrate suitable for use in fish and non-human animal diets, said oilseed comprising a heat-treated dehulled oilseed and said protein concentrate having: from about 38% to about 58% protein on a dry weight basis; from about 52% to about 77% protein on a lipid-free dry weight basis; v' -51 O from about 2.7% to 4.6% methionine and cystine calculated as a percent of protein; S- from about 4.3% to about 7.9% lysine calculated as a percent of said protein; from about 24% to 37% lipid on a dry weight basis; 00oo from about 1.7% to 10% crude fibre on a lipid-free dry weight basis reduced concentrations of water soluble antinutritional factors in an 0 amount of at least 20% by weight relative to non-treated oilseed protein. 12. The protein concentrate of claim 10, wherein said dehulled oilseed is greater than 55% dehulled. 13. The feed product of claim 7, wherein said feed having from about 40 to about 80% protein calculated on a lipid-free dry weight basis, said source being adapted for use in animal and aquafeeds whereby said admixture is characterized by at least one of the following: enriched concentrations of essential amino acids and bio-available minerals relative to those present in said animal protein source or untreated oilseed; enriched concentrations of highly unsaturated n-3 fatty acids relative to those present initially in said oilseed if said source of animal offal is fish; reduced concentrations of heat-labile and water soluble and antinutritional factors in an amount of at least 20% by weight relative to non-treated oilseed protein; increased protein digestibility relative to non-treated oilseed protein; and a lipid concentration of less than 12% of dry weight of said source. -52- 14. The product of claim 12 and 13, wherein said oilseed is selected from _o canola, rape seed, soybeans, sunflower seed, flax seed, mustard seed, O cotton seed, hempseed and mixtures thereof. 15. The product of claim 12 and 13, wherein said animal protein source is selected from whole fish, fish processing waste, fish offal, fish by-catch, squid 00 i n offal, whole birds without feathers, poultry offal, beef offal, lamb offal and mixtures thereof. CI 16. A protein and lipid-rich oilseed meal suitable for use in fish and non-human animal diets comprising a heat-treated dehulled oilseed prepared according to the process of claim 1, said oilseed meal having: from about 26% to about 40% protein on a dry weight basis; from about 48% to about 64% protein on a lipid-free dry weight basis; from about 2.4% to about 4.6% methionine and cystine calculated as a percent of said protein; from about 3.6% to about 6.1% lysine calculated as a percent of said protein; from about 21% to 52% lipid on a dry weight basis; from about 2% to about 12% crude fibre on a lipid-free dry weight basis. ,-53- 0 17. A process for preparation of nutritionally upgraded oilseed meals, which are f protein and lipid-rich and have a reduced fibre content, and plant oils from O oilseeds for use in fish or other non-human animal diets or human foods Scomprising the steps of: providing a source of oilseed; subjecting said oilseed to heat treatment to substantially oo reduce the concentration of at least some antinutritional components present in said oilseed to obtain heat-treated particulate 0 seed; providing a source of unhydrolyzed animal offal; blending said heat-treated seed in particulate form with said animal offal, and if required water together with an antioxident, to form a mixture thereof; cooking said mixture under conditions selected to substantially improve protein digestibility, and substantially free cellular water present in said animal offal, as well as to facilitate separation of protein from the lipid in said animal offal and said oilseeds to obtain a cooked mixture; and separating said cooked mixture into a stickwater fraction, a moisture containing protein-rich fraction, and an animal feed grade oil fraction. 18. The process according to claim 17, further including the step of stabilizing said plant oils by adding an antioxidant. 19. The process according to claim 17, further including the step of drying said protein-rich fraction to reduce its moisture content to below about The process according to claim 17, wherein said heat treatment is a rapid heat treatment.

21. The process according to claim 17, wherein said oilseed is selected from ,-54- 0 the group consisting of canola, rape seed, soybeans, sunflower seed, flax seed, mustard seed, cotton seed, hemp and mixtures thereof.

22. The process according to claim 17, wherein said oilseed is selected from the group consisting of canola, sunflower seed, flax seed, mustard seed and mixtures thereof.

23. The process according to claim 17, wherein said animal offal is selected oo from the group consisting of fish processing waste, whole fish, fish by-catch, squid offal, whole birds without feathers, beef offal, poultry offal, lamb offal and mixtures thereof.

24. The process according to claim 17, wherein said oilseed and said animal offal are mixed together in a ratio of about 10:90 to about 90:10 by weight. The process according to claim 17, wherein said oilseed is treated to dephytinize said oilseed.

26. The process according to claim 17, further comprising the step of extracting said protein rich fraction with a solvent.

27. The process according to claim 26, wherein said solvent includes hexane.

28. In a process for the preparation of nutritionally upgraded oilseed meal from co-processing of animal offal with oilseed for use in fish or other non-human animal feeds, wherein the process includes the steps of providing a source of oilseed and cold pressing said oilseed to substantially reduce the particle size of said oilseed to yield a high value human grade oil and protein and lipid-rich meal with reduced fibre content; the improvement comprising the further steps of: providing a source of unhydrolyzed animal offal; blending said protein and lipid-rich meal with said animal offal, and if required water together with an antioxident, to form a blended mixture thereof; cooking said blended mixture under conditions selected to O substantially improve protein digestibility, and substantially free cellular water present in said animal offal, as well as to facilitate O separation of protein from the lipid in said animal offal and said Soilseeds to obtain a cooked mixture; and separating said cooked mixture into a stickwater fraction, a moisture containing protein-rich fraction, and an animal feed grade oo oil fraction. (i~r

29. The process according to claim 28, further including the step of extracting Ssaid protein and lipid-rich meals with a solvent.

30. The process according to claim 29, wherein said solvent includes hexane.

31. The process according to claim 29, wherein said oilseed is treated to dephytinize said oilseed.

32. The process according to claim 28, said process further comprising the steps of: subjecting said oilseed to heat treatment at a temperature and time sufficient to deactivate, destroy or reduce concentration of some antinutritional factors present in said oilseed, improve digestibility and reduce moisture content; providing said source of unhydrolyzed animal product selected from the group consisting of fish processing waste, whole fish, fish by-catch, squid offal, beef offal, lamb offal and whole birds without feathers; cooking said mixture at a temperature for a time sufficient to improve protein digestibility and free the bound water present in said animal offal and facilitate the separation of protein from the lipid in said animal product and said oilseed; removing fluid comprised of lipid and water soluble components from said mixture to obtain a pressed cake; and, v -56- 0- drying the pressed cake at a temperature for a time sufficient for the pressed cake to reach a moisture content of about 7 10% to O provide a protein concentrate.

33. The process of claim 32, including the further step of condensing the stickwater.

34. The process according to claim 33, wherein said heat treatment is carried (0 out at a temperature of about 100-115oC for a time of about 1.5 to about mins. The process according to claim 33, further including the step of dehulling said oilseed.

36. The process according to claim 35, wherein said dehulling is carried out by impact or disc process coupled with a gravity screening and/or air- classification process.

37. The process according to claim 33, wherein said oilseed is selected from the group consisting of canola, rape seed, soybeans, sunflower seed, flax seed, mustard seed, cotton seed, hemp and mixtures thereof.

38. The process according to claim 37, wherein said oilseed is selected from the group consisting of canola, soybeans, sunflower seed, cotton seed and mixtures thereof.

39. The process according to claim 38, wherein said offal is fish offal. The process of claim 33, further including the step of: adding a palatability enhancer to said mixture prior to the cooking step or after the pressing step.

41. The process according to claim 32, further including the step of adding an antioxidant to the mixture prior to the cooking step or after the pressing step.

42. The process according to claim 40, wherein said palatability enhancer is selected from the group consisting of products based on krill, euphausiids, sV 57- o squid and mixtures thereof. o 43. The process according to claim 42, wherein said antioxidant is selected Sfrom the group consisting of ethoxyquin, butylated hydroxyanisole, butylated hydroxytoluene, Vitamin E and mixtures thereof.

44. In a process for preparation of nutritionally upgraded oilseed meals, which are protein and lipid-rich and have a reduced fibre content, and plant oils n from oilseeds for use in fish or other non-human animal diets or human foods the improvement comprising the steps of: providing a source of oilseed; subjecting said oilseed to a drying step to obtain oilseed having a moisture content of less than 10% to thereby improve dehulling of said oilseed; dehulling said oilseed to provide a source of dehulled oilseed; providing a source of unhydrolyzed animal offal; blending said dehulled oilseed with said animal offal, and if required water together with an antioxident, to form a mixture thereof; cooking said mixture under conditions selected to substantially improve protein digestibility, and substantially free cellular water and lipids present in said animal offal, as well as to facilitate separation of protein from the lipid in said animal offal and said oilseeds to obtain a cooked mixture; and separating said cooked mixture into a stickwater fraction, a moisture containing protein-rich fraction, and an animal feed grade oil fraction.

45. The process according to claim 44, wherein said oilseed is treated to dephytinize said oilseed.

46. The process according to claim 44, further comprising the step of extracting said protein rich fraction with a solvent. -58-

47. The process according to claim 44, wherein said solvent includes hexane. o 48. A process for preparation of nutritionally upgraded oilseed meals, which are protein and lipid-rich and have a reduced fibre content, and plant oils from oilseeds for use in fish or other non-human animal diets or human foods comprising the steps of: providing a source of oilseed; o c subjecting said oilseed to heat treatment to substantially Sreduce the concentration of at least some antinutritional components normally present in said oilseed to obtain heat-treated seed; dehulling said heat-treated seed to produce a meat fraction, a hull fraction or a mixture thereof; cold pressing said meat fraction or said mixture to yield said plant oils and said protein and lipid-rich meals; providing a source of unhydrolyzed animal offal; blending said protein and lipid-rich meal with said animal offal to form a blended mixture thereof; cooking said blended mixture under conditions selected to substantially improve protein digestibility, and substantially free cellular water present in said animal offal, as well as to facilitate separation of protein from the lipid in said animal offal and said oilseeds to obtain a cooked mixture; and, separating said cooked mixture into a stickwater fraction, a moisture containing protein-rich fraction, and an animal feed grade oil fraction. 20 October 2005 Her Majesty the Queen in the right of Canada as represented by The Minister of Fisheries and Oceans