AU4016001A - Composition and method - Google Patents

Description

Regulation 3.2

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

(ORIGINAL)

IP Australia Documents received on.

0 1 MAY 2001 Batch No: E Name of Applicant: Actual Inventors: Address for Service: Brian Rodney Quinn AND Irving Goodfellow AND Robert Francis Paton Pigott QUINN, Brian Rodney GOODFELLOW, Irving PIGOTT, Robert Francis Paton DAVIES COLLISON CAVE, Patent Attorneys, 1 Little Collins Street, Melbourne, Victoria 3000.

"Composition and method" Invention Title: Details of Associated Provisional Application No: PQ7236 2 May 2000 The following statement is a full description of this invention, including the best method of performing it known to us: Q OPER\MKR'GliNER G .\\PQ7236-O-CAP.OC l15i01 P OPER\M KR'SPECI\grow\lhap -2- COMPOSITION AND METHOD FIELD OF THE INVENTION The present invention relates to animal growth promoting compositions and related methods and uses which involve the administration of organic selenium. In particular, but not exclusively, the organic selenium is in the form of protein which includes selenium substituted amino acids or amino acid analogues, preferably combined with a chemical denaturing or proteolytic agent such as, for example, urea.

BACKGROUND OF THE INVENTION The present invention has been conceived in the context of our rapidly increasing world population with awareness of the resulting burden this brings to bear upon our natural environment in order to feed, clothe, shelter and entertain increasing numbers of people.

There is as a result a pressing need within agriculture to devise more efficient means of cultivating crops and rearing animals. The present inventors have surprisingly found o means by which growth rates of animals can be significantly increased, which in turn allows for more efficient provision of animal based products and more efficient use of agricultural resources.

oooo• S Thompson, K.M. et al ("Growth and plasma triiodothyronine concentrations are modified by selenium deficiency and repletion in second-generation selenium-deficient rats", The Journal of Nutrition, 1995, vol 125 pp 864-873) studied the reversal of growth depression in rats which had been fed a selenium deficient diet resulting from selenium administration. This document also mentions that triiodothyronine infusion to selenium deficient rats had no reversal effect upon growth depression, thereby suggesting that conversion of inactive thyroxine (T 4 to active 3,5,3-triiodothyronine (T 3 which could theoretically be catalysed by selenium administration, is not responsible for reversal of growth depression in rats.

p:1oPeR11.JMSPEClh~okg-1h doc-01,0501 -3- It was reported by Master, D.G. et al ("Marginal deficiencies of cobalt and selenium in weaner sheep: response to supplementation", Australian Journal of Experimental Agriculture, 1990, vol 30 pp 337-341) that administration of a combination of copper, cobalt and selenium had an effect on clean wool growth and live weight change in weaner wether sheep. This document suggested that administration of cobalt, copper and selenium in combination resulted in an increase in clean wool growth and that when either selenium or cobalt are administered alone there is a trend toward wool growth.

Chinese Patent No. 1101799 to Zhang discloses that a fowl fodder additive including amounts of a number of rare earth metals, including selenium sulphate, could have some positive effect on fowl growth. Japanese Patent No. 2973074 to Asahi Kasei Kogyo KK teaches that it is possible to prevent parasitic growth in fish by culturing the fish in sea water which includes a selenium containing microorganism. This document teaches that feeding the fish on the selenium containing microorganism prevents parasitic infection.

Finally, Soviet Union Patent No. 1433441 discloses a growth promoting composition for carp fish which includes iodine and selenium dissolved in a solution of gelling agent and mixed with hydrolysing yeast which has been dried and fragmented.

None of the documents referred to above disclose or suggest that the growth rate of nonselenium deficient animals of a diverse range of species can be improved by administering organic selenium to the animal. In particular, there is no disclosure or suggestion within the prior art that proteins containing selenium substituted amino acids or amino acid analogues may form the basis of an animal growth promoting composition. This is quite a :surprising outcome as prior to the present invention it was generally considered that 25 selenium, when presented in a bound form as organic selenium (or more particularly as protein or amino acids containing selenium), would not be sufficiently bioavailable to exhibit any significant physiological activity.

The present inventors have investigated the effects of feeding an extract obtained from fish, (particularly, but not exclusively from European carp (Cyprinus carpio)) upon the growth rates of a range of different animals. European carp were introduced into P:'PERXNIKRISPEChgr-ih-p do-01/O0501 -4- Australia's Murray basin waterways in the 1960s and have since reached plague proportions, causing reduced native fish numbers and severe riverbank erosion. European carp are able to out-compete other fish species and have had a devastating environmental impact by interrupting the river food chain, depleting the rivers of small crustaceans and vegetable matter and causing regular blue green algal blooms. The European carp therefore comprises a significant environmental threat to Australia's waterways, and is a pest which needs to be controlled.

Surprisingly, the present inventors have determined that an extract obtained from fish, and particularly from European carp, can be used to promote animal growth. Furthermore, when experimenting with compositions comprising fish extracts to which it was intended to raise the nitrogen level by the incorporation of urea, it was surprisingly observed that the growth rates of animals fed on such compositions were significantly and unexpectedly improved.

Without wishing to be bound by theory, the present inventors have postulated that the oooe growth promoting compositions according to the present invention are effective as a result i of conversion of inactive thyroxine (T 4 to active 3,5,3-triiodothyronine (T 3 through the oooo incorporation of selenium into the enzyme iodothyronine deiodinase. The present inventors have speculated that the significance of the presence of urea in such compositions and the effectiveness of the urea containing compositions in promoting animal growth results from denaturation of the selenium containing protein, making the selenium more readily bioavailable. Other denaturing agents or means of denaturing or even cleaving protein may therefore be used in the same way, within the context of the present invention.

S* It is an object of the present invention to provide animal growth promoting compositions and methods of promoting animal growth. Other objects of the present invention will become apparent from the following detailed description thereof.

P:1OPERWKMRSPECIr oa lh-pr doc-01/0501 SUMMARY OF THE INVENTION According to one embodiment of the present invention there is provided an animal growth promoting composition comprising organic selenium.

According to another embodiment of the invention there is provided an animal growth promoting composition comprising protein which includes selenium substituted amino acids or amino acid analogues.

According to a further embodiment of the present invention there is provided an animal growth promoting composition comprising selenium substituted amino acids or amino acid analogues.

According to another embodiment of the present invention there is provided an animal growth promoting composition comprising a fish derived protein containing extract and a physiologically acceptable chemical denaturing or proteolytic agent.

According to still a further embodiment of the invention there is provided a method of promoting growth of an animal which comprises administering organic selenium to the animal.

.00o0: In another embodiment of the invention there is provided a method of promoting growth of an animal which comprises administering to the animal protein which includes selenium substituted amino acids or amino acid analogues.

09-00 In a further embodiment of the invention there is provided a method of promoting growth of the animal which comprises administering to the animal selenium substituted amino acids or amino acid analogues.

P:WPERM KR\SPECJ-lh-c~p do-.OIOOiS -6- According to still a further embodiment of the present invention there is provided a method of promoting growth of an animal which comprises administering to the animal a fish derived protein containing extract and a physiologically acceptable chemical denaturing or proteolytic agent.

According to another embodiment of the invention there is provided use of organic selenium in preparation of a composition for administration to an animal to promote growth.

Also provided by the invention is the use of protein which comprises selenium substituted amino acids or amino acid analogues in preparation of a composition for administration to an animal to promote growth.

In another embodiment of the invention there is provided the use of protein which comprises selenium substituted amino acids or amino acid analogues and a physiologically acceptable chemical denaturing agent in preparation of a composition for administration to an animal to promote growth.

In a preferred embodiment of the invention the animal growth promoting compositions include a chemical denaturing or proteolytic agent. Preferably the chemical denaturing 20 agent is urea.

In another preferred embodiment of the invention the fish derived protein containing extract is derived from European carp (Cyprinus carpio).

According to another embodiment of the invention there is provided a method of preparing an animal growth promoting composition which comprises the steps of: •oo00 obtaining an amount of finely divided fish which is a source of protein containing selenium substituted amino acid or amino acid analogues; combining the fish with between 5%w/w and 200%w/w of water to produce an aqueous fish mixture; and P 1OPER\MKR'SPEC1\rosd,.P doc-/05101 -7heating the mixture to between about 40'C and about 100°C for between about 5 minutes and about 10 hours.

In a preferred embodiment a chemical denaturing agent, proteolytic agent and/or emulsifier is added prior to, during or after the heating step. In a further preferred aspect one or more physiologically acceptable additive, excipient and/or active ingredient is added prior to, during or after the heating step.

DETAILED DESCRIPTION OF THE INVENTION Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the common general knowledge in Australia.

20 In its very broadest aspect the present invention relates to an animal growth promoting composition comprising organic selenium and to methods involving such a composition.

By the term "organic selenium" it is intended to convey that the selenium is incorporated within an organic compound. For example, such compounds may be seleno-alkanes, seleno-alkenes, seleno-alkynes, seleno-aryl compounds and cyclic or heterocyclic selenosubstituted compounds. In particular, the selenium substituted compounds may constitute biological compounds such as selenium substituted nucleic acids, nucleosides or nucleotides or their analogues, selenium substituted proteins, amino acids or their analogues or selenium substituted carbohydrates or lipids. It is particularly preferred that the animal growth promoting compositions according to the present invention comprise protein which includes selenium substituted amino acids or amino acid analogues or the selenium substituted amino acids or amino acid analogues themselves. The term P: OPERVA KR\SPEC1\Sro%,h-cap do -8- "selenium substituted amino acid or amino acid analogue" is intended to embrace all naturally occurring or synthetic amino acids, imino acids or other amino acid or imino acid like compounds which can be incorporated within proteins or protein like polymers, and which can be substituted by selenium. Particular examples of amino acids which lend themselves to selenium substitution are seleno-cysteine or its oxidised dimeric form, cystine, and seleno-methionine. In both cases the sulphur groups of the native amino acid are substituted with a selenium atom.

In a preferred aspect of the invention the animal growth promoting composition comprises protein which includes selenium substituted amino acids or amino acid analogues.

Preferably the protein within such compositions has been denatured, by for example S 'treatment with one or more chemical denaturing agents, heat, high or low pH or X or UV radiation or cleaved by proteolytic agents such as proteases (eg pepsin, trypsin) and Smicrobial digesting agents, as will be well understood by a person skilled in the art. It is 15 possible for this denaturation or proteolysis to take place during production of the composition and (in the case of denaturation) to then store the composition under appropriate conditions for the protein to be maintained in a denatured form until the time •of administration. Preferably, however, a chemical denaturing agent or chaotropic agent or proteolytic agent may also be incorporated within the composition. In this circumstance it 20 is obviously important for the agent to be physiologically acceptable in the animal species to which the composition is to be administered, at least in the amounts of the agent which are to be incorporated within the composition. Chemical denaturing agents or chaotropic agents which can be used to denature proteins are well known in the art, examples of which include urea, guanidine hydrochloride, sodium iodide, sodium perchlorate, sodium dodecylsulfate, performic acid, P-mecaptoethanol and other agents such as for example those referred to in Anal.Biochem.121, 382-387 (1982) and DE-A-373442. In the case where a chemical denaturing agent or proteolytic agent is adopted in production of the compositions according to the invention, which is not physiologically acceptable or not physiologically acceptable in the amount used, it may be necessary for this material to be removed from the composition before administration by filtering or separation means, as well understood in the art. Preferably the chemical denaturing agent adopted is urea.

P:'OPER\IK R\SPEC\gromh-p.dcmo 11j05i0 -9- Organic selenium compounds utilised within the present invention may be synthetically produced or extracted from natural sources. In particular, means of synthetically producing selenium containing amino acids or amino acid analogues and proteins which comprise them are well known. In one preferred embodiment of the invention protein which includes selenium substituted amino acids or amino acid analogues is extracted from fish. A wide variety of fish species may be utilised in order to extract protein which includes selenium containing amino acids or amino acid analogues. For example carp, catfish, Murray cod, snapper, barramundi, whiting, brim, flathead, trout, tuna, salmon, mackerel, garfish, blue grenadier or other fish species which are a source of selenium substituted amino acids may be utilised in the present invention. It will be recognised, however, that some species of fish will contain higher levels of selenium substituted amino acids and may therefore constitute a more efficient source from which the selenium substituted amino acids or proteins comprising them can be derived. It will also be 00 15 understood that the levels of selenium substituted amino acids within particular fish populations may depend upon the age, weight and diet of the fish as well as the nature of 0: its environment and the time of year at which it is harvested. The present inventors have particularly found that European carp (Cyprinus carpio), especially that harvested from the waterways of the Australian Murray basin, includes particularly high levels of selenium 20 substituted amino acids. As European carp is a pest which is present in plague proportions in Australian inland waters it offers a source of selenium substituted amino acids and the proteins which contain them which are readily available at low economic cost, the harvest of which also provides an environmental benefit.

The present invention additionally provides methods of promoting the growth of animals involving the administration of organic selenium to an animal, the growth rate of which is intended to be increased. Animals which may be treated by the methods according to the present invention include for example agricultural animals such as cattle, sheep, goats, pigs; domestic animals such as cats, dogs, rabbits, guinea pigs; captured wild animals such as kangaroos, koalas, wombats, wallabies, lions, tigers, elephants, giraffes, deer, buffalo; reptiles such as crocodiles, snakes, lizards; birds such as chickens, turkeys, emus, P'?.PERW KRSPEC\grol-ap dc-O I05/01 ostriches, canaries, budgerigars, cockatoos, parrots; fish and other aquatic animals. The term "animal" as used herein is also intended to embrace humans.

The methods of the invention are intended to increase the rate of weight gain of an animal relative to an animal of the same species, weight, size and sex which is fed an identical diet without administration of organic selenium. The methods of the invention may additionally be adopted to prevent fatigue and to prevent development and/or growth of tumors. Preferably, the methods of the present invention involve administration of proteins comprising selenium substituted amino acids or amino acid analogues or the selenium substituted amino acid or amino acid analogues themselves. In the case where selenium containing proteins are administered it is preferable for the protein to be administered in a denatured or cleaved form. This may be achieved by denaturation or proteolysis prior to administration and may involve administering the protein in S. combination with a chemical denaturing agent or proteolytic agent, as outlined above.

15 Preferably, the chemical denaturing agent is urea.

For example, the administration may be oral or parenteral and may occur between one or five times daily, may take place on alternate days, may be once or a couple of times per week, may be once or a couple of times per fortnight or may be as little as only once or a couple of times per month. In fact, one off administrations may also be effective in promoting animal growth at least to some extent. Preferably, however, between lmg/kg bodyweight and about 500mg/kg bodyweight of selenium containing protein or selenium containing amino acid or amino acid analogue is administered to an animal per day.

Administration may be by injection, such as by intravenous, intrarterial, subdermal, intraperitoneal or intramuscular injection or alternatively administration may be enteral, for example via oral, sublingual buccal or rectal administration. In preferred forms of the invention administration is oral for example via an oral drench or by incorporation of organic selenium into animal feed or water supply.

P:OPERNI KRSPEC'grosh- p d-01/5101 -11 A further aspect of the present invention relates to the use of organic selenium in the preparation of compositions for administration to animals to promote growth. Also included within the scope of the invention is a method of preparing an animal growth promoting composition which involves obtaining an amount of finely divided fish which is a source of protein containing selenium substituted amino acids or amino acid analogues, combining the fish with water, preferably between about 5% and about 200% water (by weight of the fish component) to produce an aqueous fish mixture, and then heating the mixture. The fish utilised in this process may be any of a selection of fish species, as outlined above, and may include all or only some selected parts of the fish carcass including skin, scales, fins, bones, flesh and internal organs as well as the contents of the gastro intestinal tract. For efficiency of the extraction process the fish material will ,preferably be cut, ground, pulverised or pureed into fine pieces or preferably into a pulp.

Heating of the mixture will preferably be conducted to between about 40'C and about 100'C for between about 5 minutes and about 10 hours. It is additionally possible for a chemical denaturing agent and/or emulsifier to be added prior to, during or after the heating step. One or more physiologically acceptable additive, excipient and/or active ingredient may also be added prior to, during or after the heating step. Physiologically acceptable additives and excipients include for example colouring agents, flavouring agents, stabilisers, preservatives, buffers, anti-oxidants as well as other food additives or S 20 other materials routinely incorporated within veterinary or pharmaceutical products. By reference to "active ingredients" it is intended to mean that physiologically active agents i such as for example anthelminthic agents, antibiotic agents, fungicidal agents, growth hormones or paraciticides may also be incorporated with the compositions of the invention.

It is to stressed, however, that such materials have be mentioned here by way of example only and that these recitations are not intended to be limiting upon the scope of the invention. It is also possible for nutritional supplements such as vitamins, minerals and the like to be incorporated within compositions of the invention.

In the process of preparing compositions of the invention it is possible to isolate an insoluble protein containing phase from the heated aqueous fish mixture by way of conventional filtration or other separation techniques. It is this insoluble protein P: OP ERM K RS PEC\ro, th-cp.do -O1105101 -12containing aqueous phase which is believed to include the substantial amount of the selenium containing protein derived from the fish material. It may be desired to utilise the insoluble protein containing phase or the aqueous fish mixture in a dried form by, for example, utilising conventional evaporation or membrane filtration techniques. Although a substantial amount of the selenium containing protein will be present in the insoluble fraction, some will also be present in the aqueous phase. The aqueous phase may therefore be useful in methods of the invention especially if concentrated by mild heating.

The compositions according to the present invention can be administered in liquid form such as in the form of a liquid suspension, in the form of a paste, powder, granules or tablets by utilising techniques well known in the art for the formulation of such dosage forms.

ee e.

The present invention will now be further described by way of example only, with oo 15 reference to the following non-limiting examples;

EXAMPLES

Example 1 S 20 6 thoroughbred racehorses in training were fed a standard racing diet. 3 horses also received an organic selenium containing supplement (as prepared according to the protocol outlined in example 5) orally at the rate of 10ml per day. The group receiving the supplement performed better than the group receiving no supplementation.

Example 2 New Zealand white rabbits were weaned at 3 weeks of age and placed on a commercial rabbit pellet plus clover hay ad libitum. 25 rabbits were divided into 5 groups so that equal numbers of males and females were present in each group. Each rabbit in the trial was daily dosed orally with 0.5ml of organic selenium supplement (as prepared according to the protocol outlined in example 5) as follows. Weights were recorded at 3 weeks of age and again at 12 weeks. Results are shown in Table 1.

P:'OPER\MKR\SPECI\ row~h-cp.doc -01/05/01 -13- TABLE 1 Trial Control Snapper Murray Barramundi Catfish Carp Group> Cod Weight at 3 387 grams 423 394 442 431 440 Weeks grams grams grams grams grams Weight at 3604 3648 3694 3681 3720 4102 12 Weeks grams grams grams grams grams grams Weight 3217 3225 3300 3239 3289 3662 Gain grams grams grams grams grams grams Weight Gain Index 1 1.002 1.025 1.007 1.022 1.138 Change over Control 0.2 2.5 0.7 2.2 13.8 Example 3 In another trial, feral goat kids were dosed orally with 5ml of organic selenium supplement (as prepared according to the protocol outlined in example 5) per week as shown below for a period of 4 weeks. Initial selection into treatment groups was made so that sexes and initial bodyweights were the same for each group. Results are shown in Table 2.

o WR\SPEC1\g,.,p doc-0101 -14- TABLE 2 Trial Control Carp Group> Average Weight at Start 3.74 kg 3.90 kg Average Weight After 4 Weeks 7.38 kg 7.83 kg Weight Gain 3.64 kg 3.93 kg Average Gain in Grams Per Day 130 140 Weight Gain Index 1 1.08 Change over Control 8 0 0 0 0**0 Example 4 A total of 103 feral goats were shipped by truck on a journey lasting 8.5 hours. Of these 50 received a single oral dose of organic selenium (prepared according to example 5) one week prior to shipping. During transit 4 goats in the untreated group died whilst none in the treated group died.

Example The trial was executed using 10 Hereford steers all of around 10 months of age. They were divided randomly into two groups of 5. The trial group received 10ml of organic selenium (prepared according to the protocol below) per day whilst the controls received none. All animals were weighed at the start and end of the trial period (4 weeks). In addition all animals were tested for their Glutathione (GSHPx) peroxidase levels at the end of week 5. The results for both weight and GSHPx levels are shown in Tables 3, 4 and The steers were running on a good quality mixed pasture and received no supplementary feed. They were dosed orally once daily with the supplement.

P.'OPERMKRSPECIgrolthlap doc-01 05/01 The preparation of the organic selenium supplement involved utilising whole carp (46% by weight of end product) which was minced and mixed with a quantity of water and heated to around 65 C for 30 to 60 minutes after achieving the target temperature. The product was then pureed through an in line emulsifier. Finally urea was added to the mix and the product was stirred for a further 20 minutes.

RESULTS OF GSHPx TRIAL ON STEERS Treated Group Weight Gain TABLE 3 Steer Number Weight 26/12/99 kg Weight 25/1/00 kg Weight Gain kg 164 317 344 27 166 281 305 24 165 324 342 18 169 224 261 36 163 297 334 37 Total Gain kg 142 Average Gain kg 28.4 Average Gain Per Day in kg 0.916 0 Control Group Weight Gain TABLE 4 Steer Number Weight 26/12/99 kg Weight 25/1/00 kg Weight Gain kg C1 317 333 16 C2 294 318 24 C3 338 356 18 C4 286 305 19 274 299 Total Gain kg 102 Average Gain kg 20.4 Average Gain Per Day in kg 0.658 The results for weight gains are not statistically significant.

P:\O PERkAI R\PECIp lh-P.Am -0 1 /05101 16- Results for Glutatione Peroxidase Levels TABLE Control Group Treated Group GSHPx U/GHb GSHPx U/GHb 18 39 24 31 22 29 13 33 .100 Average 19.6 3402 Standard Deviation 4.28 4.6 T Test 0.009 The results for GSHPx determinations are statistically significant. This response is not surprising as the selenium status for all animals is deficient.

Example 6 Ten thoroughbred racehorses on standardised diet were selected and divided into two groups of five so that both groups contained horses at the same stage of training and sexes 10 were equally distributed. The standard diet consisted of concentrate present as grain and a mixture of roughages including lucerne, clover, wheaten and oaten chaff. Hay and water were available at all times. All horses received supplementation in the form of minerals (including selenium) and vitamins. The control group of horses received no organic selenium supplement whilst the treated group received 10 ml daily by mouth of organic selenium supplement prepared according to example The results of this trial are shown in Table 6.

KW!OERW M(RSPEChromh-pslpd xc0 1 10,101 17- TABLE 6 RACEHORSE TRIAL FOR T3/T4/GSHPX ESTIMATIONS.

CONTROL

HORSES CONTROL CONTROL CONTROL CONTROL T4 T3 T3/T4 GSHPX 1 12 0.63 0.053 129.000 12 0.62 0.052 132.000 6 9 0.69 0.077 151.000 7 19 0.89 0.047 112.000 9 7 0.8 0.114 120.000 AVERAGE 11.80 0.73 0.07 128.80 STDEV 4.55 0.12 0.03 14.69 TREATED TREATED TREATED TREATED TREATED HORSES T4 T3 T3/T4 GSHPX 2 16 0.95 0.059 141.000 3 17 0.97 0.057 109.000 4 17 1.02 0.060 129.000 8 10 0.86 0.086 131.000 34 0.83 0.024 119.000 AVERAGE 18.80 0.93 0.06 125.80 STDEV 8.98 0.08 0.02 12.21 STATISTICAL 0.29 0.07 0.64 0.74 ANALYSIS N/S N/S N/S N/S The results show that there were no statistically significant differences between groups which is not surprising due to the completeness of the diet and the healthy selenium status of both groups as shown by the Glutathione Peroxidase readings. A remarkable observation is that despite the lack of significance in all groups, the T3 levels border on significant which would indicate that there has been an increase in the conversion of T4 to T3 in the treated animals. This is even more surprising given that the groups are small and the overall selenium status of the horses in both groups is the same.

P:'OPERWMKRSPECgrosthlcbp dM-o0 i151i 18- Example 7 Ten dogs (Jack Russell Terriers) of mixed ages and sexes were divided into 2 groups of five so that ages and sexes were equal in both groups. All dogs were fed on a standardised diet consisting of chicken necks, minced beef and dry food. The treated group of dogs received 2ml organic selenium supplement prepared according to example 5 daily by mouth for 10 days. At the end of the trial blood samples were taken from all dogs to assess T3, T4 and GSPX labels. Results are shown in Table 7.

TABLE 7 DOG TRIAL FOR T3/T4/GSHPX ESTIMATIONS.

CONTROL CONTROL CONTROL CONTROL CONTROL DOGS T4 T3 T3/T4 GSHPX 2 25 0.71 0.028 169 3 23 0.45 0.020 135 4 35 0.91 0.026 164 30 0.95 0.032 153 9 37 0.82 0.022 138 AVERAGE 30.00 0.77 0.026 151.800 STDEV 6.08 0.20 0.005 15.156 TREATED TREATED TREATED TREATED TREATED DOGS T4 T3 T3/T4 GSHPX 1 23 0.92 0.040 153 6 25 1.02 0.041 136 7 20 1.2 0.060 150 8 23 0.99 0.043 165 10 88 1.48 0.017 131 AVERAGE 35.80 1.12 0.040 147.000 STDEV 29.24 0.23 0.015 13.657 STATISTICAL 0.64 0.04 0.09 0.40 ANALYSIS N/S P=.04 N/S N/S It was not surprising that all dogs exhibited excellent selenium status as shown by the Glutathione levels. It is remarkable given such small sample sizes that the T3 levels showed a significant difference between treated and control levels with the T3/T4 ratios bordering on significant.

Claims (23)

19- THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS: 1. An animal growth promoting composition comprising organic selenium. 2. An animal growth promoting composition comprising protein which includes selenium substituted amino acids or amino acid analogues. 3. A composition according to claim 2 wherein the protein is denatured. 4. The composition according to claim 3 wherein denaturation has been achieved using one or more chemical denaturing agents, heat, high or low pH or radiation. A composition according to claim 4 wherein the chemical denaturing agent is urea, guanidine hydrochloride, sodium iodide, sodium perchlorate, sodium dodecylsulfate, performic acid and/or p-mecaptoethanol. 6. The composition according to claim 4 wherein the chemical denaturing agent is urea. 20 7. An animal growth promoting composition comprising selenium substituted amino acids or amino acid analogues. 8. An animal growth promoting composition comprising a fish derived protein containing extract and a physiologically acceptable chemical denaturing or proteolytic agent. 9. The composition according to claim 8 wherein the protein containing extract is derived from carp, catfish, Murray cod, snapper, barramundi, whiting, brim, flathead, trout, tuna, salmon, mackerel, garfish, blue grenadier or other fish which is a source of selenium substituted amino acids. P:'OPERI\MKRSPECPVrouth-cp doc-0105101 The composition according to claim 8 wherein the protein containing extract is derived from European Carp (Cyprinus carpio). 11. The composition according to any one of claims 8 to 10 in which the chemical denaturing agent is urea. 12. The composition according to any one of claims 8 to 11 which is in a paste, powder, granule, tablet or liquid form. 13. Composition according to any one of claims 8 to 11 which is in an aqueous liquid suspension form. The composition according to any one of claims 8 to 13 further comprising one or more physiologically acceptable additive, excipient and/or active ingredient. A method of promoting growth of an animal which comprises administering organic selenium to the animal. 16. A method of promoting growth of an animal which comprises administering to the S 20 animal protein which includes selenium substituted amino acids or amino acid analogues. 17. The method according to claim 16 wherein the protein is denatured. 18. The method according to claim 17 wherein the protein is administered in conjunction with a physiologically acceptable chemical denaturing or proteolytic agent. 19. The method according to claim 18 wherein the denaturing agent is urea. P.\OPER\IKR\SPECI\growh-cap.doc-0 I/0,S01 -21 The method according to any one of claims 16 to 19 wherein the protein is derived from fish.

21. The method according to claim 20 wherein the fish is carp, catfish, Murray cod, snapper, barramundi, whiting, brim, flathead, trout, tuna, salmon, mackerel, garfish, blue grenadier or other fish.

22. The method according to claim 20 wherein the fish is European carp (Cyprinus carpio).

23. A method of promoting growth of an animal which comprises administering to the animal selenium substituted amino acids or amino acid analogues.

24. The method according to any one of claims 15 to 23 wherein the administration is oral or parenteral.

25. The method according to any one of claims 15 to 24 wherein administration is between one and five times daily.

26. The method according to any one of claims 15 to 24 wherein the administration is on alternate days.

27. The method according to any one of claims 15 to 24 wherein the administration is once weekly.

28. The method according to any one of claims 15 to 24 wherein the administration is once fortnightly.

29. The method according to any one of claims 15 to 24 wherein the administration is once monthly. P:1OPERVO KR SPEC\gr-olh-ip docOIO5I01 22 The method according to any one of claims 16 to 24 wherein a dose rate of between about 1mg/kg and about 500mg/kg of protein is administered to the animal per day.

31. Use of organic selenium in preparation of a composition for administration to an animal to promote growth.

32. Use of protein which comprises selenium substituted amino acids or amino acid analogues in preparation of a composition for administration to an animal to promote growth.

33. Use of protein which comprises selenium substituted amino acids or amino acid analogues and a physiologically acceptable chemical denaturing or proteolytic agent in preparation of a composition for administration to an animal to promote growth.

34. The use according to claim 33 wherein the chemical denaturing agent is urea.

35. The use according to either claim 33 or claim 34 wherein the protein is derived from fish.

36. The use according to claim 35 wherein the fish is European carp (Cyprinus carpio).

37. The use according to any one of claims 33 to 36 wherein the composition is in a paste, powder, granule, tablet or liquid form.

38. The use according to any one of claims 33 to 36 wherein the composition is in an aqueous liquid suspension form.

39. The use according to any one of claims 33 to 38 wherein the composition includes one or more physiologically acceptable additive, excipient and/or active ingredient. P:'OPER1M KRiSPECgro,1ih-cip doc-OI105101 -23- A method of preparing an animal growth promoting composition which comprises the steps of: obtaining an amount of finely divided fish which is a source of protein containing selenium substituted amino acids or amino acid analogues; combining the fish with between 5%w/w and 200%w/w of water to produce an aqueous fish mixture; and heating the mixture to between about 40°C and about 100°C for between about 5 minutes and about 10 hours.

41. The method according to claim 40 wherein one or more of a chemical denaturing, a proteolytic agent and an emulsifier is added prior to, during or after the heating step. 15 42. The method according to either claim 40 or claim 41 wherein one or more physiologically acceptable additive, excipient and/or active ingredient is added prior to, during or after the heating step.

43. The method according to any one of claims 40 to 42 wherein a product of step is 20 subject to separation to isolate an insoluble protein containing phase.

44. A method according to any one of claims 40 to 43 wherein a product of step is **subject to drying.

45. An animal growth promoting composition, methods of promoting growth of an animal, uses of protein in preparing compositions and methods of preparing animal growth promoting compositions substantially as hereinbefore described with reference to the examples. DATED this 1st day of May, 2001 Brian Rodney Quinn AND Irving Goodfellow AND Robert Francis Paton Pigott By DAVIES COLLISON CAVE Patent Attorneys for the Applicant