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( 12 ) United States Patent ( 10) Patent No .: US 10,813,960 B2 Stamets ( 45 ) Date of Patent : * Oct

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( 12 ) United States Patent ( 10) Patent No .: US 10,813,960 B2 Stamets ( 45 ) Date of Patent : * Oct US010813960B2 ( 12 ) United States Patent ( 10 ) Patent No .: US 10,813,960 B2 Stamets ( 45 ) Date of Patent : * Oct . 27 , 2020 ( 54 ) INTEGRATIVE FUNGAL SOLUTIONS FOR ( 56 ) References Cited PROTECTING BEES AND OVERCOMING COLONY COLLAPSE DISORDER ( CCD ) U.S. PATENT DOCUMENTS 6,106,867 A * 8/2000 Mishima A23G 3/48 ( 71 ) Applicant: Paul Edward Stamets , Shelton , WA 424/539 ( US ) 6,183,742 B1 2/2001 Kiczka 6,660,290 B1 12/2003 Stamets ( 72 ) Inventor : Paul Edward Stamets , Shelton , WA 7,122,176 B2 10/2006 Stamets ( US ) 7,951,388 B2 5/2011 Stamets 7,951,389 B2 5/2011 Stamets ( * ) Notice : Subject to any disclaimer , the term of this 8,501,207 B2 8/2013 Stamets patent is extended or adjusted under 35 8,753,656 B2 6/2014 Stamets 8,765,138 B2 7/2014 Stamets U.S.C. 154 ( b ) by 0 days. 9,399,050 B2 7/2016 Stamets This patent is subject to a terminal dis 9,474,776 B2 10/2016 Stamets claimer. 2002/0146394 A1 10/2002 Stamets 2004/0161440 A1 8/2004 Stamets 2004/0209907 A1 * 10/2004 Franklin A61K 31/517 ( 21 ) Appl . No.: 15 /332,803 514 / 266.22 2004/0213823 Al 10/2004 Stamets ( 22 ) Filed : Oct. 24 , 2016 2005/0176583 A1 8/2005 Stamets 2005/0238655 A1 10/2005 Stamets ( 65 ) Prior Publication Data 2005/0276815 Al 12/2005 Stamets US 2017/0035820 A1 Feb. 9 , 2017 2006/0171958 A1 8/2006 Stamets 2008/0005046 Al 1/2008 Stamets 2008/0046277 A1 2/2008 Stamets Related U.S. Application Data 2008/0264858 Al 10/2008 Stamets 2009/0047236 A1 2/2009 Stamets ( 63 ) Continuation - in - part of application No. 14 / 641,432 , 2009/0047237 A1 2/2009 Stamets filed on Mar. 8 , 2015 , now Pat. No. 9,474,776 , and a continuation - in - part of application No. 14 / 247,207 , (Continued ) filed on Apr. 7 , 2014 , now abandoned . FOREIGN PATENT DOCUMENTS ( 60 ) Provisional application No. 62 / 074,023 , filed on Nov. AU 2015229670 B2 2/2017 2 , 2014 , provisional application No. 61 / 967,117 , filed AU 2017200872 B2 1/2019 on Mar. 10 , 2014 . ( Continued ) ( 51 ) Int . Ci . OTHER PUBLICATIONS A61K 36/07 ( 2006.01 ) A61K 36/074 ( 2006.01 ) Revilla et al . Comparison of Several Procedures Used for the A61K 45/06 ( 2006.01 ) Extraction of Anthocynains From Red Grapes ; J. Agric . Food Chem . A23K 50/90 ( 2016.01 ) 46 , pp . 4592-4597 . ( Year: 1998 ) * A23K 20/163 ( 2016.01 ) ( Continued ) A23K 20/147 ( 2016.01 ) A23K 20/158 ( 2016.01 ) AOIN 65/00 ( 2009.01 ) Primary Examiner — Amy L Clark A23K 10/30 ( 2016.01 ) ( 74 ) Attorney, Agent, or Firm - Michael Best & A23K 20/111 ( 2016.01 ) Friedrich LLP A23K 20/10 ( 2016.01 ) A23L 33/145 ( 2016.01 ) A61K 9/12 ( 2006.01 ) ( 57 ) ABSTRACT A01K 51/00 ( 2006.01 ) ( 52 ) U.S. Cl . The present invention is based on a plurality of benefits from CPC A61K 36/07 ( 2013.01 ) ; A01K 51/00 the extracts of mycelia of individual fungal species , and ( 2013.01 ) ; A01N 65/00 ( 2013.01 ) ; A23K 10/30 mixtures of species , to provide an armamentarium of ( 2016.05 ) ; A23K 20/10 ( 2016.05 ) ; A23K defenses from multiple stressors in order to help bees 20/111 ( 2016.05 ) ; A23K 20/147 ( 2016.05 ) ; survive a complex of symptoms collectively called colony A23K 20/158 ( 2016.05 ) ; A23K 20/163 collapse disorder ( CCD ) . More particularly, the present ( 2016.05 ) ; A23K 50/90 ( 2016.05 ) ; A23L invention utilizes specific concentrations of extracts from 33/145 ( 2016.08 ) ; A61K 9/12 ( 2013.01 ) ; A61K pure cultured mycelium from mushroom forming fungi to 36/074 ( 2013.01 ) ; A61K 45/06 ( 2013.01 ) ; reduce harmful viruses in bees and to increase the longevity A23V 2002/00 ( 2013.01 ) of bees . ( 58 ) Field of Classification Search CPC A61K 36/074 ; A23K 50/90 See application file for complete search history . 30 Claims , 7 Drawing Sheets US 10,813,960 B2 Page 2 ( 56 ) References Cited OTHER PUBLICATIONS U.S. PATENT DOCUMENTS Kusum Harjai et al . Pathogens and Disease , vol . 69 , Issue 1 , Oct. 2009/0130138 A1 5/2009 Stamets 2013 , pp . 62-65 . ( Year: 2013 ) . * 2010/0086647 A1 4/2010 Kristiansen Stamets, P. Mycelium Running: How Mushrooms can help save the 2011/0008384 A1 1/2011 Stamets World . 2005. tenspeedpress: New York . p . 281. ( Year: 2005 ) . * 2011/0200551 A1 8/2011 Stamets Entomopathogenic Fungus, Wikipedia article accessed via https :// 2012/0039976 A1 2/2012 Stamets en.wikipedia.org/wiki/Entomopathogenic_fungus on Aug. 13 , 2015 . 2012/0070414 Al 3/2012 Stamets Jarosz - Wilkolazka et al . , “ Oxalate Production by Wood -Rotting 2013/0183418 A1 7/2013 Maurice Fungi Growing in Toxic Metal - Amended Medium , ” Chemosphere , 2013/0287829 A1 10/2013 Stamets 2014/0065131 A1 3/2014 Kelly vol . 52 , ( 2003 ) , pp . 541-547 . 2014/0065263 A1 * 3/2014 Kelly A23L 31/00 MSDS for Oxalic Acid ( ScienceLab.com , created Oct. 10 , 2005 , 426/60 updated May 21 , 2013 , pp . 1-6 ) . 2014/0105928 A1 4/2014 Stamets Stamets, “ Growing Gourmet and Medicinal Mushrooms, ” Ten Speed 2014/0220150 Al 8/2014 Stamets Press, Berkeley, California , ( 1993 ) , pp . 42-43 . 2015/0335689 Al 11/2015 Stamets Stamets , “ Myco Medicinals : An Information Treatise on Mush 2016/0000754 A1 1/2016 Stamets rooms, ” 3rd Ed . Commonly asked Questions , MycoMedia Produc tions , Olympia , Washington , ( 1999 ) , p . 65 . FOREIGN PATENT DOCUMENTS Holland and Barrett, “ Product Information : Unbeelievable Health CA 2257840 A1 * 12/1997 AOIN 37/02 Bee Prepared Max Strength , ” Apr. 2014 created date , p . 1 ( http :// CA 2939143 C 7/2019 www.hollandandbarrett.com/pages/product_detail.asp?pid=4729 ) . CN 1432291 A 7/2003 Revilla et al . , “ Comparison of Several Procedures Used for the 101524142 A 9/2009 Extraction of Anthocynains from Red Grapes , ” J. Agric . Food 102100810 A 6/2011 Chem . , vol . 46 ( 1998 ) , pp . 4592-4597 . CN 102366028 A 3/2012 Stamets, P.E., et al ., “ Extracts of Polypore Mushroom Mycelia CN 102771786 A 11/2012 Reduce Viruses in Honey Bees ,” Nature Scientific Reports 8 : 13936 EA 032759 B1 7/2019 EP 3110260 B1 1/2018 ( 2018 ) | DOI : 10.1038 / s41598-018-32194-8 . JP 60186260 A 9/1985 Espacenet Machine Translation of JP2011184349 ( A ) Sep. 22 , 2011 JP 63020384 A 1/1988 ( prepared by EPO Espacenet on Sep. 25 , 2019 ) . JP 63020384 A2 1/1988 New Zealand First Examination Report for Application No. 723135 JP 06343479 A * 12/1994 dated Feb. 19 , 2020 ( 4 pages ) . JP 07000144 A 1/1995 Pylywpi et al . , “ Sodium Benzoate and Potassium Sorbate Preser JP H08154627 A 6/1996 vatives in Juices and Fruits, ” The Connecticut Agricultural Experi JP 11169107 A 6/1999 JP 11169107 A2 6/1999 ment Station , New Haven , Bulletin 921 ( 1994 ) . JP 2006083112 A 3/2006 New Zealand Patent Office Examination Report No. 2 for Appli JP 2010163458 A * 7/2010 cation No. 723135 dated Jun . 26 , 2020 ( 5 pages ) . JP 2011184349 A 9/2011 Ibrahim et al . , “ Effect of artificial culture media on germination , JP 2011184349 Al 9/2011 growth , virulence and surface properties of the antomopathogenic KR 1020110098071 A 9/2011 hyphomycete Metarhizium anisopliae, ” Mycological Research , Jun . WO 2002028189 4/2002 2002 , 106 ( 6 ) : 705-15 . WO 2002065836 A2 8/2002 WO 2005067955 A1 7/2005 Carreck et al . , “ Honey bees can disseminate a microbial control WO 2006121350 A1 11/2006 agent to more than one inflorescence pest of oilseed rape ,” Biocontrol WO WO08097482 A2 8/2008 Science and Technology , Mar. 2007 , 17 ( 2 ) : 179-91 . WO 2012018266 A1 2/2012 WO 2015138361 A1 9/2015 * cited by examiner U.S. Patent Oct. 27 , 2020 Sheet 1 of 7 US 10,813,960 B2 100 LEGEND 90 CONTROL I. obl . 0.1 % 80 I. obl . 1 % 704 I. obl . 10 % 60 50 SurvivalPercent 40 30 20 104 1 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 Time ( Days ) FIG . 1 100 LEGEND 90 CONTROL 80 G. res. 0.1 % G. res. 1 % 70 G. res . 10 % 60 50 SurvivalPercent 40 30 20 104 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 Time ( Days ) FIG . 2 U.S. Patent Oct. 27 , 2020 Sheet 2 of 7 US 10,813,960 B2 LEGEND CONTROL 0.1%F.pin. %1pin.F. 10%F.pin. 181920212291011121314151617 Time(Days) FIG.3 123456789 100 90 80 70 60 50 40 30 20 10 0 Survival Percent U.S. Patent Oct. 27 , 2020 Sheet 3 of 7 US 10,813,960 B2 LEGEND Control F.fom.1% 1'2'3'4'5'6'7'8'9101112131415161718192021222324252627 Time(Days) FIG.4 100 80 60 40 20 0 Survival Percent U.S. Patent Oct. 27 , 2020 Sheet 4 of 7 US 10,813,960 B2 30 LEGEND CONTROL F.fom.0.1% F.fom.1% F.fom.10% 25 20 1 15 Time(Days) FIG.5 10 5 0 1.0 0.8 0.6 0.4 0.2— 0.C Surviving U.S. Patent Oct. 27 , 2020 Sheet 5 of 7 US 10,813,960 B2 8,000 I. obliquus 7,000 Week 2 6,000 5,000 Week 1 TotalVirusParticles 4,000 3,000 7 2,000 1,000 :: 0 Time 0 Control 0.10 % 1 % 10 % Control 0.10 % 1 % 10 % FIG . 6 25,000 G. resinaceum 20,000 Week 1 Week 2 15,000 TotalVirusParticles 10,000 5000 | 0 Time 0 Control 0.1 0 % 1 % 10 % Control 0.10 % 1 % 10 % FIG . 7 U.S. Patent Oct. 27 , 2020 Sheet 6 of 7 US 10,813,960 B2 2,500 F.
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