CA2420266A1 - Oral omega-3 fatty acid neutritional supplement for companion animals - Google Patents

Abstract

A powdered or microencapsulated nutritional additive for companion animals, for use as top dressing or the supplementation of commercially available pet and other companion animal foods, to increase serum levels of EPA and DHA in companion animals, derived from fish oil and/or seal oil, the use of a powdered microencapsulated nutritional additive derived from fish oil and/or seal oil to increase serum levels of EPA and DHA
in companion animals; and the use of a powdered microencapsulated nutritional additive derived from fish oil and/or seal oil in prevention or treatment of renal, dermatological and cardiovascular disease or as an anti-inflammatory, and in the prevention or treatment of conditions associated therewith, in companion animals.

Description

~) i s c ~ o s~.~-...v~ . ; -~ ~ . .",. : ~ , ~ .'. ..
L i ~~: ~,_ ~:::________~ __~ ~ . __.~-iL~__ ._~ _~ __. _.- -._ Ir'_':=;:en~.«iW i.~~;:vl~
r~.:ei~.,.._ . ~I~~ r!,;~ ar,r_ 1,:~a~;:,.
i.,...''!W;i t%'i :=!-~l i~W _",!.'im,.... .:: _:1_ ~i"' . '~~;~:t:!CI iii:
i.li~ ~~~~' L~l:~t:Ci ,'_~~'l~m.._~iL .:_ . _.. ~,. v_u. __v .~..L~!.~_._._.~. Li~ ~l~,i,"~; t'"y :_~4m~~~~-.:
1 . t r' ~ ,' ~ r y l_ _:. ' r ~' ~ ~~1 __...._......_..._ :C; '.-\,..u.__..v.. ~_ .. Y, __u.-... _. ..._ ,~. _..v.
.._1~ ' _ . .. ~ .~ v.w___ .u. .~

FOR COMPANION ANIMALS
Field of the Invention The present invention relates to a nutritional supplement for companion animals rich in the Omega-3 Fatty Acids, and particularly rich in Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA). More particularly, the present invention relates to a powdered omega-3 fatty acid rich supplement for companion animals such as dogs, cats and the like, prepared from fish and/or seal oil.
Background of the Invention Fodder and livestock nutritional additives for swine, livestock and the like are known, particularly for the purpose of enhancing the nutritional value of the food produced from such animals, such as meat and dairy products,. United States Patent No.
5,106,639 relates to a dried fatty fodder additive containing omega-3 fatty acids and a process for producing same, to produce meat with minimal off taste characteristics particularly associated with such additives. U.S. Patent No. 4,971,820 relates to a process for the production of a pellet type animal feed containing edible liquid lipids.
Fish oil extracts have been used in the preparation of animal feed products.
U.S. Patent no. 4,284,652 relates to a process for a matrix like pet food which may comprise fish oils. U.S. Patent No. 5,693,358 and International Application No.
PCTIAU00/00129, published August 31, 2000 as WO 00/49889 relate to further examples of processes for the preparation of animal feed supplements and the feed supplements made therefrom for use with livestock to improve the nutritional qualities of the food products made therefrom while reducing the taint usually associated with excessive use of fish oils.
The positive health effects of omega-3 fatty acids, particularly in association with fish and/or seal oil, has been well documented, including in PCT/PT01/00020, published August 16, 2001 as WO 02/13838 A1.

There is however a need for a nutritional supplement for companion animals such as dogs, cats and other like mammals to provide the positive health affects for such animals associated with omega-3 fatty acids, which are associated with products originating from fish and/or seal oil. There is in particular a need for such products which are convenient to use, which retain their freshness and avoid off or tainted tastes, in the relatively small quantities which are associated with companion animals.
Summary of the Invention The present invention provides for a powdered or microencapsulated nutritional additive for companion animals, for use as top dressing or the supplementation of commercially available pet and other companion animal foods, to increase serum levels of EPA and DHA in companion animals, derived from fish oil and/or seal oil.
The present invention further provides for the use of a powdered microencapsulated nutritional additive derived from fish oil and/or seal oil to increase serum levels of EPA
and DHA in companion animals.
The present invention further provide for the use of a powdered microencapsulated nutritional additive derived from fish oil and/or seal oil in prevention or treatment of renal, dermatological and cardiovascular disease or as an anti-inflammatory, and in the prevention or treatment of conditions associated therewith, in companion animals.
Detailed Description of the Preferred Embodiment omega-3 tatty acids are dietary essential fatty acids (EFAs) that play an important role in a variety of physiological functions including neurological development, inflammation and membrane-mediated functions (1). Essential Fatty Acids (EFAs) in general are dietary factors whose ingestion is essential for physiological functions. They were discovered in 1929 by George and Mildred Burr (University of Minnesota, USA).
There are two series of EFAs, the n-6 (or omega-6), derived of cis-linoleic acid;
and the n-3 (or omega-3), derived of alfa-linolenic acid. The digits indicate the position of the first double link (C=C) in the molecule counted from the omega terminal. As for certain vitamins, cis-linoleic and alfa-linolenic acids are devoid of activity (except that of serving as energy substrates - being "burnt" in cellular metabolism to generate energy

-2-for the cell) while they have not been specifically bio-transformed in the human body, as shown hereafter.

-3-Series n-6 Series n-3 cis-linoleic acid alfa-linolenic acid ~4 ~ delta-6-desaturase (D6D) gams-linolenic acid (AGL) 18:4, n-3 y elongation y dihomo-gamma-linolenic acid (DGLA) 20;4:, n-3 y 1 delta-5-desaturaser. (DSD) y y PG's series ~ arachidonic acid eiceosa-pentaenoic acid (20:5, n-3) y y y y y y ~ PG's series 3 PG's series 2 longer EFAs longer EFAs The EFAs of these two series (n-6 and n-3) are not interchangeable in animals.
It is worthy noting the low activity of delta-5-desaturase no Man and the guinea-pig, while it is high in mice and rats.
EFAs are important for two fundamental reasons: (a) they are constituents of all membranes in all tissues of the body, playing a role which is vital for determining the characteristics and the biological capacities of their respective membranes -it is therefore logic that EFAs deficiencies trigger great dysfunctions in all body tissues;
and (b) EFAs are the precursors of a group of highly reactive and very efemerous substances - prostaglandins (PG's), leuco-trienes (LT's) and other related molecules, such .as thromboxanes (TX's), which intervene in an infinity of cell and organ processes. More than 50 PG's, LT's and related molecules with biological importance have been identified, acting at local messengers in the regulation of the activity of different tissues and organs where they are produced. Arachidonic acid (AA) has very different and varied effects through the products it generates. Thus, for example, it both originates PG12 (prostaciclin), with desirable effects of inhitition of platelet aggregation and vasodilatation, and thromboxane A2 (TXA2) and PGF2a, which promote vasospasm, thrombosis and inflammation. After AA, PG's and thromboxane.
(TX's) are due to the intervention of a ciclo-oxigenase, while LT's are die to a completely different enzine - a lipo-oxigenase:

-4-AA in membrane deposit (inhibited by NSAI's and PGEI) free AA
10~
(inhibited by vit and and bt Lipooxigenase Ciclooxigenase (inhibited by NSAI's and AAS) a OH-acid derived from DGLA) LTs PG's and TXA2 (After Horrrobin, 7982) Seal Oil is known to be particularly rich with omega-3 EFAs. It has been shown that EFAs are essential to many metabolic processes including synthesis and maintenance of cellular membranes structures. The body is incapable of synthesizing EFAs and when they are not present in sufficient quantity, the body replaces them with the most similar Fatty Acids (FA) it can find in its existing reserves. Such deficiency states normally involve serious cellular dysfunctions, affecting different organs, not only because the EFAs intervene directly in metabolic chains as substrates or regulators (an example of that being the metabolism of prostaglandins), but also because deficiency alters significantly the permeability and the selectivity of different cellular membranes, thus excessively impeding and/or facilitating the passage of substances across cell membranes. This causes clinical symptoms related with deficiency and/or excess of metabolites and other products which are detrimental for the good functioning of cells.
When this occurs, organs which are important for bodily equilibrium tend to suffer, as, for example, happens with the central nervous system (CNS), the peripheral nerves, the blood vessels, the immune system, the joints, the mucosal membranes, i.e.
ocular, nasal, intestinal, genito-urinary, etc. and the skin. Resulting dysfunctions are related to both omega-3 EFA deficiencies, and excesses of certain non-essential FA which act as omega-3 EFA antagonists. Therefore, one of the consequences found with prolonged and intense Omege-3 EFA deficiencies is the alteration of cell membrane functions and structure. Frequent consequences includes: (a) edematous fluid collections -very often "of lymphatic type", and (b) the different types of the "inadequate secretion of

-5-anti-diuretic hormone syndrome", usually associated with "lymphatic edema" and "orange skin cellulitis".
Fish oils have docosa-pentaenoic acid (DPA) concentrations only about 1/10 of those of EPA and DHA. Therefore fish oil supplements do not have great impact on circulating DPA concentrations, and any impact observed probably was mediated by EPA
elongation to DPA. Benistant et al. (1996) found that DPA can accumulate in endothelial cells phospholipids and reduce A4'--s undesirable effects; and Kanayasu-Toyoda et al. (1996) found that DPA has no effect upon vascular smooth muscle cells migration but that it can, (trough, phospholipidic-DPA) increase 20 fold (dose-dependently) the migration of vasculares endohtelial cells, while Hogdson et al. (1993) demonstrated that DPA concentration human blood platelets sanguineas is inversely correlated with the rate of deposition of those platelets in the coronary arteries of women (it should be remembered that there are 10 times more deaths vascular causes among women than those due to breast cancer).
Several studies suggest clearly that DHA may be the object of a shortening of its chain so as to generate EPA, while EPA can be easily converted to DPA.
Once Kanayasu-Toyoda et al. (1996) and Benistant et al. (1996) and many other authors demonstrated that EPA, DHA and DPA induce significant effects upon the regulation of circulation and of diverse cellular behaviours, it was easy to understand that these EFAs, present in fish and seal oil (particularly DPA), participate in the fine regulation of those mechanisms which help maintain health, preventing or curing coronary disease, thrombosis, vascular inflammatory processes, the production of tri-glycosides and of low density lipoproteins (LDL) and other adverse cardio-vascular affects. It was also demonstrated that seal fat and DPA in particular are capable of reducing the conversion of AA to thromboxane A2 (TXA2, a thrombogenic agent) and to enhancer production of to prostaglandin D3 (PGD3, a platelet-anti-aggregation agent), a series of effects which can only be favorable to health and the fight against disease.
On the other hand, it has recently been recognized that natural oils (namely those of vegetable origin), if extracted under heat (between 160 and 200°C) and high pressure, are low or even devoid of, omega-3 EFAs, as a consequence of omega-3 EFA
stereo transformation, through the rotation of asymmetrical carbons, i.e. passing from cis (or -~6-cis-cis) form to traps (or cis-traps) form, as is the case, for example, of cis-gamma-linolenic acid, which at temperatures oscillating between 54 and 60°C
transforms itself into traps-gamma-linolenic acid. Almost all alimentary vegetable fats available today are precisely extracted under high heat (160 to 300°C) and high pressure of circa 2,5 to 3 atmospheres, resulting in much higher yields of those fats. Such newly formed stereo-isomers are metabolically antagonists to any remaining omega-3 EFAs, acting as isosteric andlor allosteric competing antagonists.
Seal Oil is endowed with a specific EFA - DPA, or Docosa-pentaenoic acid (22:5, n-3) which has been shown to induce (a) inhibition of vascular smooth muscle cells;
and (b) the migration of endothelial cells and the endothelial repair of blood vessels; an effect confirming its usefulness in the prevention of vascular diseases, namely, that of arteriosclerosis; it has been shown that DPA is far more active than other EFAs; and that other EFAs, specifically those which exist in fish-oils, such as EPA (eicosa-pentaenoic acid [20:5,n-3]) and DHA (docosa-hexaenoic acid [22:6,n-3J), recognized as useful in the tight against vascular diseases, act through their transformation into DPA, which has been shown to be 10 times more active than EPA in the migration of endothelial cells.
That explains the greater potency of seal oil and its derivatives containing DPA.
Fish oils of course are an important source of omega-3 EFAs although seal oil is particularly well endowed in all EFAs essential to mammals.
Fish oil components of particular interest are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Supplementation with omega-3 fatty acids has been shown to be beneficial for the treatment of renal, dermatological and cardiovascular disease in dogs (2,3,4). Administration of diets with high levels of omega-3 fatty acids has been found to be renoprotective in experimentally induce renal failure in dogs when compared with diets high in omega-6 fatty acids or saturated fats (2,5). omega-3 fatty acid administration has also been demonstrated to slow the progression and spread of lymphoma in dogs. Dogs with lymphoma were found to have a significantly higher disease-free interval and survival time when fish oil was added to the diet as compared with standard chemotherapy alone (6).
Administration of EPA and DHA may affect inflammatory processes by altering the body's response to injury. Cells react to insult by releasing the eicosanoid precursors, including arachidonic acid (AA) and EPA, from the cell membrane, thereby activating the cyclooxygenase (CO) and lipoxygenase (LO) enzyme systems to produce the eicosanoids (7). The type of eicosanoid produced is dependent on the amount of each fatty acid available and the activity of the respective enzyme systems (8).
The relative concentrations of various omega-3 and omega-6 fatty acids in the diet are reflected in the membrane phospholipid composition of inflammatory cells and the subsequent eicosanoids produced upon their release (9). Normally AA is the primary substrate and is metabolized by cyclooxygenase to form the proinflammatory 2-series prostaglandins and thromboxane A2, a potent vasoconstrictor and promoter of platelet aggregation (10).
The action of lipoxygenase on AA results in the release of the leukotriene-4 group of compounds, which are also proinflammatory. Conversely, when EPA acts as the substrate, the 3-series prostaglandins and thromboxanes and 5-series leukotrienes that are produced are anti-inflammatory and only weakly vasoconstrictive (7). These changes are the result of the competition between AA and EPA or DHA for various metabolic pathways. In this way, EPA and DHA may be useful components in the treatment of a variety of conditions in dogs and cats.
Small amounts of DHA can be obtained from eggs and some meats (11) but EPA and DHA are present in largest amounts in fish and fish by-products such as meal or oil (12).
Alpha-linolenic acid, another omega-3 fatty acid, serves as the precursor for EPA and DHA and can be obtained from a variety of dietary sources including flax (12).
However, the bioconversion of alfa-linolenic acid to EPA and DHA in dogs and cats is very poor and the increase in serum EPA and DHA levels following consumption of flax is negligible (8). Therefore, in order to obtain the health benefits of EPA and DHA, they must be acquired directly by consumption of fish or fishmeal in the diet or addition of a dietary supplement containing EPA and DHA. Dietary fatty acids are incorporated into phospholipid in the liver and integrated into biological membranes. Serum phospholipid levels have been demonstrated to serve as reliable biomarkers for omega fatty acid bioavailability in the diet (13).
Example A study was conducted, designed to evaluate changes in serum EPA and DHA
levels following administration of a powdered, microencapsulated omega-3 fatty acid supplement to healthy dogs and cats. The supplement was conveniently packaged in foil packages for stability and freshness.
_g_ Materials and methods Ten healthy adult dogs and 7 healthy adult cats were enrolled. Animals were privately owned, were clinically normal and were not receiving any supplements containing omega-3 fatty acids. Dogs were fed a commercial pet food (Hill's Science Diet Canine Adult Maintenance, Hill's Pet Nutrition, Topeka, KS) and dog biscuits (lams Original Formula Small Biscuits, The lams Company, Dayton, OH). Cats were fed a commercial pet food only. (Hill's Science Diet Feline Adult Maintenance, Hill's Pet Nutrition, Topeka, KS). None of the products contained fish or supplemental EPA or DHA, however the canine diet did supply a small amount of omega-3 fatty acid in the form of alfa-linolenic acid derived from flax. Other diets and treats were not permitted during the study period. Following a 14 d acclimatization period, a serum sample was collected and frozen at -700C. A microencapsulated, powdered supplement containing EPA
(182mg/g) and DHA (135 mg/g) was administered at a dose of 25mg/kg BW PO for dogs and 150mglcat PO. The supplement was administered as a top-dressing once daily for 14d. This dose provided approximately 14 mg/kg EPA and 11 mg/kg DHA. Serum samples were collected on the final day of supplementation and following a 14d washout period. Serum phospholipid profiles, measured as a percentage of total serum phospholipid, were determined via gas liquid chromatography (GLC) as has been previously described (14).
Data obtained from the dogs was analyzed using a repeated measures analysis of variance (ANOVA) and the Tukey-Kramer multiple comparisons test. Feline data was analyzed using a one-way ANOVA and Bonferroni's multiple comparisons test. A P
value of < 0.05 was considered significant for all comparisons. A statistical software package was used (GraphPad InStat, GraphPad Software Inc, San Diego, Ca).
Results Four male and 6 female dogs ranging in age from 1 to 12 y (mean 5.1 ~ 1.3 y) were enrolled. Dogs ranged in weight from 11 to 45 kg (mean 31.2 ~ 3.0 kg). Two male and female cats ranging in age from 1 to 8 y (mean 4.6 ~ 1.2 y) were enrolled. All cats were of similar size and body condition and were estimated to weigh 6 kg. One cat was lost to follow up so a final serum sample was not obtained. All animals consumed the supplement readily and no adverse effects were reported. The fatty acid compositions of the basal rations are outlined in Table 1. There were significant increases in EPA, _g_ DHA, total omega-3 fatty acid levels and omega-3: omega-6 ratio (P < 0.005) and significant decreases in AA to EPA ratios (P < 0.001 ) following oral supplementation in both dogs and cats (Tables 3 and 4 and Graphs 1 to 8). There were significant increases in the EPA metabolite DPA (C22:5n3) (P < 0.05) and significant decreases in total omega-6 fatty acid levels (P < 0.001 ) and in C22:5n6 (P < 0.05), a metabolite of AA, in dogs only. No significant change in AA levels occurred. These parameters normalized following cessation of administration of the supplement and were not significantly different from baseline values following the washout period (P <
0.05).
Discussion Demonstration of incorporation of EPA and DHA into serum phospholipid following oral supplementation is an important step in developing an effective treatment regimen in dogs and cats. It is believed that the addition of omega-3 fatty acids to the diet, particularly EPA and DHA, may alter the production of the eicosanoids resulting in a decrease in inflammation due to the production of less potent inflammatory mediators.
This study demonstrated that oral administration of a microencapsulated, powdered supplement containing EPA and DHA at a total dose of 25 mg/kg/day results in an increase in serum EPA and DHA levels and a decrease in the AA:EPA ratio in the serum phospholipid of dogs and cats. This increase in the percentage of less inflammatory omega-3 fatty acids in the serum phospholipid profile in place of pro-inflammatory omega-6 fatty acids suggests a possible therapeutic role for supplementation with EPA
and DHA.
The diets provided 3.84 kcal/g and 4.07 kcal/g for dogs and cats, respectively. For a 30 kg dog, total daily caloric intake is estimated to be 1500 kcal (Hand) and the canine diet would supply 33.75 mg EPA and DHA per day (11.25 mg EPA, 22.5 mg DHA). During the supplementation period this was increased to 783.75 mg per day (341.25 mg EPA, 442.5 mg DHA) through administration of the microencapsulated powder composed of 182 mg/g EPA and 135 mg/g DHA. The daily caloric intake for a 6 kg cat is estimated to be 326 kcal (Hand) and the feline diet would supply 16 mg EPA and DHA per day (8 mg EPA, 8 mg DHA). Supplementation increased the intake to 166 mg per day (74 mg EPA
and 92 mg DHA).
In this study, the changes produced in the omega-3 and omega-6 fatty acid levels during supplementation were no longer evident following the 14d washout period. It is therefore apparent that ongoing administration of EPA and DHA is necessary for continued benefits.
It will be apparent to a skilled person in the art that, although only certain embodiments of the invention have been described herein, modifications and variations may be made thereto, to achieve the object of the invention, within the spirit and scope of the invention broadly described herein.

FATTY ACID Gm1100 gm 14:0 0.085 14:1 0.011 15:0 0.010 16:0 2.515 16:1 0.420 18:0 0.936 18:1 4.520 18:2n6 3.203 18:3n6 0.010 18:3n3 0.407 18:4n3 0.001 20:0 0.038 20:1 0.073 20:2n6 0.036 20:3n6 0.014 20:4n6 0.052 20:3n3 0.003 20:4n3 0.000 20:5n3 0.003 22:0 0.022 22:1 0.051 22:2n6 0.000 22:4n6 0.014 22:5n6 0.001 22:5n3 0.004 22:6n3 0.006 24:0 0.002 24:1 0.010 TOTAL 12.450 SATURATED 3.609 MONOUNSATURATED 5.086 POLYUNSATURATED 3.756 TOTAL 12.450 OMEGA-6 3.332 OMEGA-3 0.424 TOTAL 3.756 Table 1: Fatty acid composition of the canine diet.

Fatty acid Day 0 a Dar 14 Day 28 C14 0.120.01 0.120.01 0.120.01 C14:1 0.100.01_ 0.100.01 0.100.01 C15:0 _ 0.100.01 0.11 0.01 0.11 0.01 ~

C16:0 16.020.57 15.230.30 15.330.40 C16:1 5 _ 0.290.03 0.31 0.04 0.340.0 ~

C18:0 _ 26.86 0.77 27.10 0.82 _ 26.35 0.95 C18:1 8.690.42 8.700.44 8.650.40 C18:2n6 19.11 0.88 18.00 0.69 18.44 0.8_4 C18:3n6 0.04 0.00 _ _0.04 0.01 0.0410.01__ C18:3n3 0.29 0.03 0.29 0.02 0.25 0.02 C18:4n3 0.00 0.00 0.00 0.00 0.00 0.00 C20:0 0.20 0.01 0.22 _0.02 0.19 t 0.02_ C20:1 0.16 0.02 0.16 0.02 0.14 0.02 C20:2n6 0.33 0.02a 0.39 0.02b 0.33 0.01 a ~

C20:3n6 1.250.10 1.190.10 1.250.09 C20:4n6 21.01 0.94 20.47 0.78 21.65 0.79 C20:3n3 0.01 0.00 0.02 _0.01 0.01 0.01 C20:4n3 0.01 0.00 0.02 0.00 0.01 0.00 C20:5n3 0.29 0.06a 0.96 0.16b 0.37 0.09a i C22:0 0.17 0.04 ,_~ I 0.14 0.03_- 0.12 0.03 C22:1 0.05 0.02 0.05 0.06 0.02 0.03 C22:2n6 0.02 0.01 0.01 0.01 0.01 0.01 C22:4n6 .23 0.16a 0.81 0.09b 0.98 0.10_ C22:5n6 _ 0.10 0.03b 0.13 0.03a 0.18 0.03a C22:5n3 1.96 0.16a 2.39 0.22b 2.05 0.12 C22:6n3 1.29 0.26x_ X2.43 0.22b 1.54 0.22a _ ~

C24:0 0.170.04 0.150.04 0.150.03 C24:1 0.70 0.10 0.70 0.09 0.60 0.07 Total n3 0.39a 6.11 0.45b 4.22 0.30a 3.84 Total n6 _ 41.00 0.06b 42.82 0.36a 43.17 0.50a n3:n6 0.090.01a _ 0.150.01b 0.01 0.01a AA/EPA 92.30 12.11 27.86 5.33b 77.09 10.13a a _ EPA + DHA 1.58 0.31 a 3.40 0.36b 1.91 0.28a Within rows, different superscripts indicate P <0.05 Table 2: Effect of administration of an omega-3 fatty acid supplement on serum phospholipid profiles in dogs (n=10).

Fatty acid Day 00 Day 14 D_ ay 28 ~ 0.1510.01 ____ 0_.120.01 0.140.01 _ 0.14 t 0.01 0.11 0.01 0.15 0._02 _ ____ ~
C 14:1 ~

C15:0 0.180.02 0.140.02 0.19 0.03_ C16:0 15.830.98 _ 15.060.40 15.160.65 C16:1 0.370.03 0.300.02 0.330.05 C18:0 27.61 t 0.70 28.04 0.66 29.93 1.16 ~ _ C18:1 12.01 0.37 11.2_20.50 12.230.52 C18:2n6 _ 23.41 0.90 22.85 1.93_ 25.25 0.66 ~

C18:3n6 0.04 t 0.01 0.02 0.01 0.01 0.01_ C18:3n3 0.220.01 0.200.01 0.21 0.02 C18:4n3 0.030.0 _0.01 0.01 0.01 0.01 C20:0 _ 0.590.02 0.680_.07__ 0.630.02 C20:1 0.39 0.0 0.32 0.02a 0.46 t 0.02b 4 _ C20:2n6 _ 0.58 0.03a 0.79 0.03b 0.68 t 0.0_4 _ C20:3n6 1.05 0.09 0.96 0.03 1.14 t 0.06 C20:4n6 10.50 0.88 _11.24 0.45 11.73 t 0.66 _ ' _ 0.02 0.01 0.01 0.01 0.03 t 0.01 C20:3n3 C20:4n3 0.04 t 0.01 0.04 0.01 0.03 t 0.02 C20:5n3 0.41 _0. 2.51 0.45b 0.30 t 0.05a 13a _ C22:0 _ 0.20 0.06 0.31 0.08 0.32 t 0.01 C22:1 0.170.04 _ 0.150.02 0.200.05 C22:2n6 0.03 0.03 0.01 0.00 0.01 0.01 C22:4n6 0.57 _0.10 0.53 0.05 0.67 0.15_ C22:5n6 0.07 0.01 a 0.07 0.02a 0.13 0.01 b C22:5n3 0.29 0.10 0.30 0.14 0.53 ~ 0.04 C22:6n3 1 1_0._21 a___ ; 2.75 0.22b 1.57 t 0.17a 1.5 C24:0 _ 0 37 0.05 0.4210.08 0.46 0.13 C24:1 1.021 0.75 0.06 0.77 t 0.08 0.
38 _____ Total n3 _ 5.84 0.51 b 2.67 0.27a Total n6 _ 36.82 0.57 37.37 1.40 2.52 t 0.39a _ 38.20 1.64 n3:n6 0.06 t 0._01 0.16 0.02b 0.07 0.01 a AA/EPA a 5.45 0.99b 43.13 t 4.51 34.89 6.03a a ' EPA + DHA 1.92 0.32a 5.26 0.57b 1.83 ~ 0.22a Within rows, different superscripts indicate P <0.05 Table 3: Effect of administration of an omega-3 fatty acid supplement on serum phospholipid profiles in cats (n=7).

Effect of oral supplementation of 1 serum EPA levels in dogs (n=9) _ . . . _ ..
_ _ ...._ _ .,.. _ .

.

~ 0 . , j ~t N ,.
Y

a 4 a~~ ~ z ~ _-~
0 ~~.

. x ~' a ~
_ I

~~ '' f '' w 0 . '' period Time ___ ___.
_ ___._ _.._.-_ -_ ___GRAPH 4 ._ -__ . ____ ._ _...,~~_ ._,._._ .. .__.._.. ' ___ ...._ _.. .__.___.._____ .. _ __ . ___ __-._ ~ Effect of oral supplementation _...___ _ .___ ' on Effect of oral supplementation on ' serum EPA level ~ serum DHA levels in cats in cats (n=7) (n=7) w 3 . 3 ~ x ,, ~ -,~, i .
.- ~ , I
~ x ~
< ~ ~ x y ~, ' ~
'"
' ~' 3 y ~ ~ 3 z G.
' ~ ~ a ~z ' ~ , "
s .; r ~

s 2.5 , ~ ~ ,~ k o s =~ ~~;~ ~ ~
~, ~ ~ 3 ~-.~.,~ a, ~ C"
a ~ ~t ~ -. Y

F ..Yf ~~ ~
"~k ~ ~~ ~
N ~
~~ , ~, ~ _~ ~
~ ~ f ~ f ~ 4!

1 F ~ ~ 1 5 s . 5 ~
, ~
~
t ~
=

~ s Q .
N
x~ ~
:"
~ E ~
~

'~ ~
, 0 ~
, ~
~
, ~ ~ "x ,e~
~~.~~~
"~ ~

1 , 1 .
"

~ , ~ ~ . ~-Q 0 ~ ~ ~~ ~. ~ t, Q 0.5 5 x~~ ~r ~
x ~. =~ ~ n~~ i 0 o 1 2 3 ; 1 2 3 Time period ~I Time period .._-- _______~_-___ ._._~ -._.____._i ,-___. __._~_ .-._- _..__ _ __ _ __ _G_RA_P H 3 __ _ ___ _ i Effect ofyoral supplementation on i combined EPA and DHA levels in cats (n=7) _- _ Q .~ 6 x I Q~ 4 Q = 2 ~ 0 Time period i _ _ _ _. __. . _... ___., Effect of oral supplementation of ', serum DHA levels in dogs (n=9) _--._..

~

_ _.

.

1 x q,=,~

,.

p0 Time period Effect of omega-3 fatty acid supplementation on combined EPAIDHA level in healthy dogs (mean ~ SE) 4 . ------,.

~ a . A. ~ '' e& : r.

a3 yF
5 r "' ' . ~a. a ~~ >
d - a ~s~
~

, x c3 .' .x, w ~~ .~2 1 ~,' ~'~ '1k"~ -~ ~ .~~, .t,..
'~ ~.

25 x ,x , i z f ~ a~ .~:
s5,~ , ' ~~ r.3~
~
' r ' ' F
~' 2 v tr x w p P~i ~
a ~ ~
k k ..~ , , ru, a T
..
s ~p ~ F~~ ' "
~
~

1 r~~
n ah .
.
r_., Day 0 Day 14 Day 28 Time Effect of omega-3 fatty acid supplementation on omega-6 fatty acid levels in healthy dogs .~-Q 44 '0 43 Time Day 0 Day 14 Day 28 Effect of omega-3 fatty acid supplementation on total omega-3 levels in healthy dogs __~___.~-..
..
__ ~.
~

~
. , r ---_x-~k ,~
~ ~ ~~ .~ ~~''w ~
~
~.
f ~t ' .. -0 ~
~
.~.
~

o r x '~ v F
~
' ' ~
~

; ~, ' 3 ~, s ;..,,.~
~
~4x : r~.,...

". ~# .
~ ~ ;z Rk ~s ~a ~;

a ~~~~

'.as .

, ,.~
x:
&
s a ~
,;

Day 0 Day 14 Day 28 Time EPA
Day 0/Day 14: P<0.01 Day 14/Day 28: P<p.01 Day O/Day 28: P=0.234 DHA
Day 0/Day 14: P<0.01 Day 14/Day 28: P<0.01 Day 0/Day 28: P=0.286 18:2n-6 Day 0/Day 14: P=0.058 Day 14/Day 28: P=0.343 Day 0/Day 28: P=0.124 20:4n-6 Day 0/Day 14: P=0.477 Day 14/Day 28: P=0.155 Day 0/Day 28: P=0.286 Total n6 Day 0/Day 14: P<0.01 Day 14/Day 28: P<0.01 Day O,~Day 28: P<0.889 _18-Reference List 1. Uauy R, Peirano P, Hoffman D, Mena P, Birch D, Birch E. Role of essential fatty acids in the function of the developing nervous system. Lipids 1996;31:S167-S176.
2. Brown S A, Brown C A, Crowell W A, Barsanti J A, Kang C, Allen Tet al.
Effects of dietary polyunsaturated fatty acid supplementation in early renal insufficiency in dogs.
J Lab Clin Med 2000;135:275-286.
3. Bond R, Lloyd DH. Combined treatment with concentrated essential fatty acids and prednisolone in the management of canine atopy. Veterinary Record 1994;134:30-32.
4. Freeman LM, Rush JE, Kehayias JJ, Ross JN, Meydani SN, Brown DJet al.
Nutritional alterations and the effect of fish oil supplementation in dogs with heart failure.
J Vet Intern Med 1998;12:440-448.
5. Brown SA, Brown CA, Crowell WA, Barsanti JA, Allen T, Cowell Cet al.
Beneficial effects of chronic administration of dietary n-3 polyunsaturated fatty acids in dogs with renal insufficiency. J Clin Lab Med 1998;131;447-455.

6. Ogilvie GK, Fettman MJ, Mallinckrodt CH, Walton JA, Hansen RA, Davenport DJet al. Effect of fish oil, arginine, and doxorubicin chemotherapy on remission and survival time for dogs wtih lymphoma. Cancer 2000;88:1916-1928.

7. Plotnick AN. The role of omega-3 fatty acids in renal disorders. J Am Vet Med Assoc 1996;209:906-910.

8. De Caterina R , Endres S, Kristensen S D, Schmidt E B. n-3 fatty acids and renal diseases. Am J Kidney Dis 1994;24:397-41 5.

9. Vaughn DM, Reinhart GA, Swaim SF, Lauten SD, Garner CA, Boudreaux MKet al. Evaluation of effects of dietary n-6 to n-3 fatty acid ratios on leukotriene B synthesis in dog skin and neutrophils. Veterinary Dermatology 1994;5:163-173.

10. Hickman M A. Interventional nutrition for gastrointestinal disease. Clin Tech Small Anim Pract 1998;13:211-216.

11. Conquer JA, Holub B. Effect of supplementation with different doses of DHA
on the levels of circulating DHA as non-esterified fatty acid in subjects of Asian Indian background. Journal of Lipid Research 1998;39:286-292.

12. Leaf A, Kang JX, Xiao Y-F, Billman GE, Voskuyl RA. The antiarrhythmic and anticonvulsant effects of dietary n-3 fatty acids. J Membrane Biol 1999;172:1-11.

13. Kobayashi M, Sasaki S, Kawabata T, Hasegawa K, Akabane M, Tsugane S.
Single measurement of serum phospholipid fatty acid as a biomarker of specific fatty acid intake in middle-aged Japanese men. Eur J Clin Nutr 2001;55:643-650.

14. Harvey RG. Management of feline miliary dermatitis by supplementing the diet with essential fatty acids. Veterinary Record 1991;128:326-329.

15. Lechowski R, Sawosz E, Klucinski UV. The effect of the addition of oil preparation with increased content of n-3 fatty acids on serum lipid profile and clinical condition of cats with miliary dermatitis. J Vet Med A 1998;45:417-424.

Claims (19)

Claims

1. ~A nutritional supplement for companion animals comprising a microencapsulated powder derived from fish oil or seal oil or fish and seal oil comprising EPA and DHA, for increase of serum levels of EPA and DHA in said companion animals.

2. ~The nutritional supplement of claim 1 comprising approximately 182 mg/g EPA and 135 mg/g DHA.

3. ~The use of a nutritional supplement of claim 1 or 2 to increase serum levels of DHA and EPA in companion animals.

4. ~The use of a nutritional supplement of claim 1 or 2 for prevention or treatment of renal disease, dermatological disease or cardiovascular disease in companion animals.

5. ~The use of a nutritional supplement of claim 1 or 2 for prevention or treatment of conditions associated with cardiovascular diseases, renal disease or dermatological disease in a companion animal.

6. ~The use of a nutritional supplement of claim 1 or 2 as an anti-inflammatory in companion animals.

7. ~The use of a nutritional supplement of claim 1 or 2 in the prevention or treatment of conditions associated with inflammation in companion animals.

8. ~The nutritional supplement of claim 1 or 2 where the companion animal is a dog.

9. ~The nutritional supplement of claim 1 or 2 where the companion animal is a cat.

10. ~The use of any one of claims 3 to 7, where the companion animal is a dog.

11. ~The use of any one of claims 3 to 7, where the companion animal is a cat.

12. ~The use of any one of claims 3 to 7, where the companion animal is a dog, in a dose of about 25 mg/kg/day body weight of EPA and DHA.

13. ~The use of any one of claims 3 to 7, where the companion animal is a cat, in a daily dose of 150 mg of EPA and DHA.

14. ~The use of any one of claims 3 to 7, where the companion animal is a dog, in a dose of about 14 mg EPA/kg/day and 11 mg DHA/kg/day.

15. ~The use of any one of claims 3 to 7 where the companion animal is a cat, in a daily dose of about 68 mg EPA/day and 84 mg DHA/day.

16. ~The nutritional supplement of any one of claims 1, 2, 8 and 9, where the nutritional supplement is administered by top dressing.

17. ~The use of any one of claims 3 to 7 and 10 to 15, where the nutritional supplement is administered by top dressing.

18. ~The nutritional supplement of any one of claims 1, 2, 8 and 9 where the nutritional supplement is in a foil package.

19. ~The use of any one of claims 3 to 7 and 10 to 15, where the nutritional supplement is in a foil package.