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(63) Continuation Inspart of Application No. PCT RE"SE SEN"I", "ES"E"NE

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(63) Continuation Inspart of Application No. PCT RE US 2010.0086647A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0086647 A1 Kristiansen (43) Pub. Date: Apr. 8, 2010 (54) FEED OR FOOD PRODUCTS COMPRISING filed on Jan. 25, 2006, provisional application No. FUNGALMATERAL 60/690,496, filed on Jun. 15, 2005. (75) Inventor: Bjorn Kristiansen, Frederikstad (30) Foreign Application Priority Data (NO) May 13, 2005 (DK) ........................... PA 2005 00710 Correspondence Address: Jun. 15, 2005 (DK). ... PA 2005 OO88O BROWDY AND NEIMARK, P.L.L.C. Jul. 15, 2005 (DK) ....................... PCTFDKO5/OO498 624 NINTH STREET, NW Jan. 25, 2006 (DK)........................... PA 2006 OO117 SUTE 300 WASHINGTON, DC 20001-5303 (US) Publication Classification 51) Int. Cl. (73)73) AssigneeA : MEDMUSHAS(DK) s HORSHOLM ( A2.3L I/28 (2006.01) A23K L/18 (2006.01) (21) Appl. No.: 11/914,318 A23K L/6 (2006.01) CI2P 19/04 (2006.01) (22) PCT Filed: May 11, 2006 AOIK 6L/00 (2006.01) (86). PCT NO. PCT/DKO6/OO2S3 (52) U.S. Cl. ................................ 426/62: 426/2: 119/230 S371 (c)(1) (57) ABSTRACT (2), (4) Date: Dec. 1, 2009 The present invention relates to feed and food compositions comprising material obtained by fermenting fungi of the Related U.S. Application Data Basidiomycetes family in a liquid medium. Interestingly, (63) DK2005/000498,continuation inspart filed onof Jul.application 15, 2005. No. PCT enhanceRE"SE Survival SEN"I",and/or support "ES"E"NE growth of normal, healthy (60) Provisional application No. 60/690,496, filed on Jun. animals. Furthermore, the compounds may modulate the 15, 2005, provisional application No. 60/761,744, microbial population in the digestive tract after oral intake. Patent Application Publication Apr. 8, 2010 Sheet 1 of 4 US 2010/0086647 A1 Fig. 1 Feed 2 (0.02 mg) D. Feed 1 (control) Patent Application Publication Apr. 8, 2010 Sheet 2 of 4 US 2010/0086647 A1 Fig. 2 is 14 Feed 4 (2 mg) Feed 3 (0.2 mg) Feed 2 (0.02 mg) Feed 1 (control) | |– |– (*):- We — Week???? 1-2 Week 1-3 Week 1-4– Patent Application Publication Apr. 8, 2010 Sheet 3 of 4 US 2010/0086647 A1 Fig. 3 E. co K2 s culture time (h) Patent Application Publication Apr. 8, 2010 Sheet 4 of 4 US 2010/0086647 A1 Fig. 4 Bioactive agent 150 --ARC-5 --Kelly 100 ... v. T. s -- C-26 i -- FENIX E 50 2 US 2010/0O86647 A1 Apr. 8, 2010 FEED OR FOOD PRODUCTS COMPRISING merged fermentation of Lentinus edodes provides no growth FUNGALMATERAL advantage to laboratory animals. 0007. The patent application WO03/020944 describes a number of methods of purifying extracellular immunostimu 0001 All patent and non-patent references cited in the lating compounds from fungi grown in liquid culture. present application, are hereby incorporated by reference in 0008 Hatvani et al., describes a method of purifying a their entirety. compound, which may be Lenthionine from the culture medium of Lentinus edodes grown in liquid culture. The FIELD OF INVENTION document describes that the purified compound in vitro has antibacterial and anticandida effect against B. megaterium, S. 0002 The present invention relates to feed and food com staphylococcus, S. pyogenes and C. albicans. It is postulated positions comprising material obtained by fermenting fungi that it has no effect on a number of other microorganisms of the Basidiomycetes family in a liquid medium. Interest including E. coli and C. jejuni. ingly, Basidiomycetes grown in a liquid culture produce com 0009 Low survival rates of fish bred in fish farms is a pounds which, when used in feed or food compositions major problem. In particular, it has proven to be excessively enhance Survival and/or support growth of normal, healthy difficult to breed white fish, such as cod, in fish farms. How animals. Furthermore, the compounds may modulate the ever, other fish, Such as salmon and trout, which are routinely microbial population in the digestive tract after oral intake. bred in fish farms also have a high lethal rate. The same applies to shrimps and prawns bred in farms. BACKGROUND OF INVENTION 0010 For producers of farm animals, it is frequently desir 0003. It is known that health-promoting effects are attrib able to obtain the largest possible weight gain in animals. Also uted to glucans from fungi and yeasts. “Shiitake fungus lethality may be a problem. (Lentinus edodes) has been attributed effects which can be 0011 Health or well-being may be influenced by the exploited for many medicinal purposes such as immuno microbial population in the digestive tract. stimulation, anti-virus, anti-tumour, etc. Studies of lentinan 0012. Accordingly, there is a need for provision of feed have shown that it stimulates the immune system of the host and food products which may improve health, modulate the in a variety of ways, Such as activation of T helper cells, microbial population in the digestive tract, improve Survival increased production of Interleukin 1 and Interleukin 2, rates of animals and/or improve growth. increased antibody production in various forms of cancer, and decreasing the cholesterol level in the blood. (Herbs for SUMMARY OF INVENTION Health, January/February, 1997: K. Jones, “Shiitake: Medi 0013. One object of the present invention is to provide feed cine in a mushroom, p. 40-50, 54; Anticancer Res, Vol. and food products, which may improve Survival. 17(4A), 1997; H. Matsouka, “Lentinan potentiates immunity 0014) Another object of the invention is to provide feed and prolongs the survival time of some patients', p. 2751 and food products, which may improve growth. 2755; Adv Appl Microbiol, Vol. 39, 1993; S. C. Jong, 00.15 Yet another object of the invention is to provide feed “Medicinal and therapeutic value of the shiitake mushroom', and food products, which may modulate the microbial popu p. 153-184, Int J Immunopharmacol, Vol. 14, 1992: K. lation in the digestive tract. Irinoda, “Stimulation of microbiocidal host defense mecha 0016. In another aspect of the invention, a food product is nism against aerosol influenza virus infection by lentinan, p. provided which is a functional food, preferably either for 971-977. Jpn. J Cancer Res, Vol. 76(1), 1985; D. Herlyn, improving or maintaining health. Said functional food is pref "Monoclonal antibody-dependent murine macrophage-me erably Suitable for consumption by human beings. diated cytotoxicity against human tumors is stimulated by 0017. The above-mentioned feed and food products com lentinan, p. 37-42). prise a bioactive agent such as a Survival enhancing agent. 0004 One active ingredient of Lentinus edodes is termed lentinan, a polysaccharide based compound described as a FIGURE LEGENDS beta-(1,3) glucan backbone with beta-(1,6) side chains. 0005. “Solid-state' reactors are routinely used for cultur (0018 FIG. 1: positive effect of different amounts of the ing fungi such as Lentinus edodes. bioactive agent in feed on the weight increase of NOROC 0006. This is a technology which is used for many pur piglets. Feeds with 0.02, 0.2 and 2.0 mg bioactive agent/kg poses such as composting, production of biological products feed (see Table 4) were given to weaned piglets and feed Such as enzymes, soy sauce, acetic acid, and the like. For the without the bioactive agent was used as control. The weight production of lentinan, Lentinus edodes can be cultivated on increase of each animal was measured weekly and the aver a suitable solid matrix provided by stems of tree or chips of age weight increase was calculated (see Table 5). The experi wood to which is often added chemical compounds Support ment is described in detail in Example 5b. ing the growth of mycelium and development of the fruiting (0019 FIG. 2: positive effect of different amounts of the bodies, where most of the lentinan is localised. The fruiting bioactive agent in feed on the feed factor of NOROC piglets. bodies are harvested, either by hand or mechanically, and are Feeds with 0.02, 0.2 and 2.0 mg bioactive agent/kg feed (see Subsequently dried and ground to a powder which can be used Table 4) were given to weaned piglets and feed without the as it is, or used in tablets, or sent for further processing Such bioactive agent was used as control. The feed factor was as extraction of lentinan. It has been Suggested that extracts of determined for weeks 1, 1-2. 1-3 and 1-4 (see Table 6). The mushrooms may be a growth promoter when fed to chickens experiment is described in detail in Example 5b. (Guo et al., British Poultry Science, 2004, 45:684-694). How 0020 FIG. 3: bacteriostatic effect of different dilutions ever, Lobanoket al., 2003, Applied Biochemistry and Micro (1:10, 1:20 and 1:40) of the bioactive agent obtained by the biology, 39:60-64, have described that mycelium from sub method as described in example 1 on E. coli K12. A culture US 2010/0O86647 A1 Apr. 8, 2010 without the bioactive agent was used as control (Ref). The whole cell fermentation culture comprising both biomass and experiment is described in detail in Example 6. extracellular growth medium, said whole cell culture com 0021 FIG. 4: cancer-cell specific cytotoxicity of different prising said bioactive agent. concentrations of LentineX—comprising an embodiment of Bioactive agents comprising a Survival enhancing activity: the bioactive agent of the present invention—on 4 different Survival enhancement is an important parameter when e.g. human and mouse cancer cell lines. The MRC-5 cell line from treating an individual for a disease, or when rearing domestic normal human fetal lung fibroblasts was used as control.
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