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(12) United States Patent (10) Patent No.: US 9.474,776 B2 Stamets (45) Date of Patent: Oct

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(12) United States Patent (10) Patent No.: US 9.474,776 B2 Stamets (45) Date of Patent: Oct US009474776B2 (12) United States Patent (10) Patent No.: US 9.474,776 B2 Stamets (45) Date of Patent: Oct. 25, 2016 (54) INTEGRATIVE FUNGAL SOLUTIONS FOR (52) U.S. Cl. PROTECTING BEES CPC ............... A61K 36/07 (2013.01); A0IK 51/00 (2013.01); A0IN 65/00 (2013.01); A23K 10/30 (71) Applicant: Paul Edward Stamets. Shelton, WA (2016.05); A23K 20/00 (2016.05); A23K 20/10 (US) (2016.05); A23K 20/III (2016.05); A23K (72) Inventor: Paul Edward Stamets, Shelton, WA 20/163A. it. (2016.05); 20. (2013.01); A23K 50/90 T.R. (2016.05); % (US) (2013.01); A61K 45/06 (2013.01); A23V - 2002/00 (2013.01) (*) Notice: Subject to any disclaimer, the term of this (58) Field of Classification Search past l S. adjusted under 35 CPC ... A01K 51/00: A01K 53/00; A23K 1/1873; .S.C. 154(b) by 0 days. A23K 50/90; A61K 36/07; A61K 36/074; AO1N 65/OO (21) Appl. No.: 14/641,432 See application file for complete search history. (22) Filed: Mar. 8, 2015 (56) References Cited (65) Prior Publication Data U.S. PATENT DOCUMENTS US 2015/0335689 A1 Nov. 26, 2015 M 6,183,742 B 1 2/2001 Kiczka Related U.S. Application Data 2005633 R 658, SN. AoN 6304 (60) Continuation-in-part of application No. 14/247.207, alleS . 424,935 filed on Apr. 7, 2014, now abandoned, which is a 2012/0039976 A1 2/2012 Stamets ................. AON 63/04 continuation-in-part of application No. 13/986,978, 424/411 filed on Jun. 20, 2013, which is a continuation-in-part of application No. 13/066,566, filed on Apr. 18, 2011, FOREIGN PATENT DOCUMENTS now Pat. No. 8,501,207, which is a division of NZ WO 2006/121350 B1 11, 2006 application No. 12/288,535, filed on Oct. 20, 2008, NZ WO 2012,01826.6 A1 2/2012 now Pat. No. 7,951,389, which is a division of k . application No. 10/853,059, filed on May 24, 2004, cited by examiner now abandoned, which is a division of application No. 09/969,456, filed on Oct. 1, 2001, now Pat. No. Primary Examiner — Kortney L. Klinkel 7,122,176, which is a continuation-in-part of (74) Attorney, Agent, or Firm — William R. Hyde application No. 09/678,141, filed on Oct. 4, 2000, now Pat. No. 6,660,290. (57) ABSTRACT (60) ySR2.5.d y N.i The present invention is based on a plurality of benefits from on Mar. 10, 2014 s u. s the extracts of mycelia of individual fungal species, and • ws mixtures of species, to provide an armamentarium of (51) Int. Cl. defenses from multiple stressors in order to help bees A6 IK 36/07 (2006.01) Survive a complex of symptoms collectively called colony A6 IK 36/074 (2006.01) collapse disorder (CCD). More particularly, the present A6 IK 45/06 (2006.01) invention utilizes specific concentrations of extracts from A2.3L I/30 (2006.01) pure cultured mycelium from mushroom forming fungi to AIK 5L/00 (2006.01) reduce harmful viruses in bees and to increase the longevity AOIK 47/06 (2006.01) of bees. AOIN 65/00 (2009.01) AOI M 1/20 (2006.01) 14 Claims, 8 Drawing Sheets 100 LEGEND as TRL ... ... F, pln, 0.1% --- F, pln, 1% ---- F, pin, 10% 5O 2 3 4 5 3 7 8 9 0 1 12 13 14 is 16 F 8 192021 22 Time (Days) U.S. Patent Oct. 25, 2016 Sheet 1 of 8 US 9,474,776 B2 1OO LEGEND 90 - CONTROL I. obl. 0.1% 80 - - - I. Obl. 1% 70 ---- I. Obl. 10% 60 50 40 30 20 10 X vs. 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 Time (Days) FIG. 1 1OO LEGEND 90 'S c- CONTROL o ooooo G, reS, 0.1% - - - G. res, 1% 70 ---- Gres, 10% 60 50 40 30 20 10 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 Time (Days) FIG. 2 US 9,474,776 B2 eAWunS Ueole U.S. Patent Oct. 25, 2016 Sheet 3 of 8 US 9,474,776 B2 | 00|| 08 09 07 eMIAunSuede U.S. Patent Oct. 25, 2016 Sheet 4 of 8 US 9,474,776 B2 CINES)ET TO?||NOO- fuAIAJnS U.S. Patent Oct. 25, 2016 Sheet S of 8 US 9,474,776 B2 8,000 7,000 6,OOO 5,000 4,000 2,000 1,000 Time O COntrol O.10% 196 109 COntrol O. 10%. 19 10% FIG. 6 25,000 G. resinaCeum 20,000 15,000 10,000 Time O Control 0.10% 1% 10%. Control 0.10% 1% 10% FIG. 7 U.S. Patent Oct. 25, 2016 Sheet 6 of 8 US 9,474,776 B2 2,500 2, O O O 1,500 1,000 500 Time O Control O.1% 1% 10% FIG. 8 1,800 1,600 1,400 1,200 1,000 800 600 400 200 Time O Control O.10% 1% 10% FIG. 9 U.S. Patent US 9,474,776 B2 pIOUSejulejoko U.S. Patent US 9,474,776 B2 pIOUSeule)KO US 9,474,776 B2 1. 2 INTEGRATIVE FUNGAL SOLUTIONS FOR insects. The annual monetary value of pollination services in PROTECTING BEES global agriculture could be as high as S200 billion. Protect ing the Pollinators, Food and Agriculture Organization of CROSS-REFERENCE TO RELATED the United Nations. The co-evolution of plants and bees APPLICATIONS (Apis species) is fundamental to their mutual Survival. The bees spread pollen and many plants produce rich nectar in This application claims the benefit of U.S. provisional return. patent application No. 62/074,023, filed Nov. 2, 2014, herein Approximately 4,000 bee species are native to North incorporated by reference in its entirety. This application is America. With the introduction of European (or “western’) also a continuation-in-part of U.S. patent application Ser. 10 honey bees (Apis mellifera) to North America by colonists, No. 14/247.207, filed Apr. 7, 2014, which claims the benefit commercial orchards and farms that would not normally be of U.S. provisional patent application No. 61/967,117, filed able to survive have thrived, although many New World Mar. 10, 2014, both of which are hereby incorporated by crops and native flowering plants are primarily dependent reference in their entirety. This application is also a con upon native bee species for pollination. Asian agriculture is tinuation-in-part of U.S. patent application Ser. No. 13/986, 15 similarly dependent upon the Asian (or “eastern') honeybee 978, filed Jun. 20, 2013, which is a continuation-in-part of (Apis cerana), although typically on a smaller and more U.S. patent application Ser. No. 13/066,566, filed Apr. 18. regionalized scale (A. mellifera has also been introduced). 2011 (now issued as U.S. Pat. No. 8,501,207), which is a divisional of U.S. patent application Ser. No. 12/288,535, Throughout agriculture the number of fruit, nut and Veg filed Oct. 20, 2008 (now issued as U.S. Pat. No. 7,951,389), etable crops benefitting from bee pollination is staggering, which is a divisional of U.S. patent application Ser. No. as are the number of flowering trees, shrubs and wildflowers. 10/853,059, filed May 24, 2004, which is a divisional of U.S. Indeed it is difficult to overstate the role of bees in the patent application Ser. No. 09/969,456, filed Oct. 1, 2001 commercial production of food. The loss of bees we are (now issued as U.S. Pat. No. 7,122,176), which is a con experiencing now is unprecedented and a huge threat to food tinuation-in-part of U.S. patent application Ser. No. 09/678, 25 security worldwide. In some regions of China, for instance, 141, filed Oct. 4, 2000 (now issued as U.S. Pat. No. the loss of bees has necessitated hand pollination to save 6,660,290), all of which are hereby incorporated by refer crops, a dauntingly difficult task. ence in their entirety. A honey bee hive is a warm, moist, densely populated environment inhabited by closely related individuals—the STATEMENT REGARDING FEDERALLY 30 perfect setting for viruses, bacteria, fungi, protozoa and SPONSORED RESEARCH OR DEVELOPMENT mites. Bees have successfully protected themselves for millions of years from such threats with unique colony-level Not Applicable. and individual-level host defense systems and immune responses, but these defenses may be breaking down as the THE NAMES OF PARTIES TO AJOINT 35 result of intense domestication of the European honey bee RESEARCH AGREEMENT and multiple threats, including new anthropogenic stressors, resulting in a precipitous decline in the number of feral Not Applicable. honey bees and native bees in areas including North America, Europe and China from 1972 to 2006, and the INCORPORATION-BY-REFERENCE OF 40 emergence of colony collapse disorder (“CCD) in honey MATERIAL SUBMITTED ON A COMPACT bees in 2006. DISC The domestic honeybee industry is dependent upon queen breeding, the process of selection that brings about the lines Not Applicable. to be propagated, and queen rearing, the process of produc 45 ing and culling queen honey bees. The large majority of bee BACKGROUND OF THE INVENTION breeding in the United States is carried out by 10-15 large queen-producing companies, who exchange genetic infor 1. Field of the Invention mation from about 500 breeder queens.
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