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(12) Patent Application Publication (10) Pub. No.: US 2017/0196922 A1 Embree Et Al US 20170196922A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2017/0196922 A1 Embree et al. (43) Pub. Date: Jul. 13, 2017 (54) METHODS FOR IMPROVING MILK Publication Classification PRODUCTION BY ADMINISTRATION OF (51) Int. Cl. MICROBAL CONSORTA A636/064 (2006.01) A23K 40/35 (2006.01) (71) Applicant: Ascus Biosciences, Inc., San Diego, A23K 50/10 (2006.01) CA (US) A6IR 9/00 (2006.01) A6II 35/742 (2006.01) (72) Inventors: Mallory Embree, Vista, CA (US); (52) U.S. Cl. Luke Picking, San Diego, CA (US); CPC .......... A61K 36/064 (2013.01); A61K 9/0053 Grant Gogul, Cardiff, CA (US); Janna (2013.01); A61K 9/0019 (2013.01); A61 K Tarasova, San Diego, CA (US) 35/742 (2013.01); A23K 50/10 (2016.05); A23K 40/35 (2016.05) (21) Appl. No.: 15/400,484 (57) ABSTRACT The disclosure relates to isolated microorganisms—includ ing novel strains of the microorganisms—microbial consor (22) Filed: Jan. 6, 2017 tia, and compositions comprising the same. Furthermore, the disclosure teaches methods of utilizing the described micro organisms, microbial consortia, and compositions compris Related U.S. Application Data ing the same, in methods for modulating the production and (60) Provisional application No. 62/276,142, filed on Jan. yield of milk and milk components in ruminants. In particu 7, 2016, provisional application No. 62/276,531, filed lar aspects, the disclosure provides methods of increasing on Jan. 8, 2016, provisional application No. 62/334, desirable components of milk in ruminants. Furthermore, the 816, filed on May 11, 2016, provisional application disclosure provides for methods of modulating the rumen No. 62/415,908, filed on Nov. 1, 2016. microbiome. Patent Application Publication Jul. 13, 2017. Sheet 1 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 2 of 21 US 2017/O196922 A1 3x&&&&&&&&&&&&&&&&&&&&&&&&& Patent Application Publication Jul. 13, 2017. Sheet 3 of 21 US 2017/O196922 A1 pe sees sees spp. xppe spot see pop box ope, ope b. e. ex sex 3.8 x: x- E. S. Patent Application Publication Jul. 13, 2017. Sheet 4 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 5 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 6 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 7 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 8 of 21 US 2017/O196922 A1 Patent Application Publication Jul. 13, 2017. Sheet 9 of 21 US 2017/O196922 A1 -{}- 3.JOSOA §§§?§§§§¿?§§§§§{{}}:$?? Patent Application Publication Jul. 13, 2017. Sheet 10 of 21 US 2017/0196922 A1 s s s 8.IOOS OW Patent Application Publication Jul. 13, 2017. 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Sheet 21 of 21 US 2017/0196922 A1 US 2017/O 196922 A1 Jul. 13, 2017 METHODS FOR IMPROVING MILK selective breeding; however, the need for more efficient PRODUCTION BY ADMINISTRATION OF production of milk and milk components per animal is MICROBAL CONSORTA required. 0006 Identifying compositions and methods for sustain CROSS-REFERENCE TO RELATED ably increasing milk production and modulating milk com APPLICATIONS ponents of interest while balancing animal health and well being have become imperative to satisfy the needs of every 0001. This application claims the benefit of priority to day humans in an expanding population. Increasing the U.S. Provisional Application No. 62/276,142, filed Jan. 7, worldwide production of milk and further modulating desir 2016; U.S. Provisional Application No. 62/276,531, filed able milk components by Scaling up the total number of Jan. 8, 2016; U.S. Provisional Application No. 62/334,816, livestock on dairy farms would not only be economically filed May 11, 2016; and U.S. Provisional Application No. infeasible for many parts of the world, but would further 62/415,908, filed Nov. 1, 2016; each of which is herein result in negative environmental consequences. incorporated by reference in its entirety. 0007 Thus, meeting global milk and milk component yield expectations, by simply scaling up current high-input FIELD agricultural systems utilized in most of the developed world is simply not feasible. 0002 The present disclosure relates to isolated and bio 0008. There is therefore an urgent need in the art for logically pure microorganisms that have applications, inter improved methods of increasing milk production and further alia, in dairy production. The disclosed microorganisms can increasing yield of desirable milk components. be utilized in their isolated and biologically pure States, as well as being formulated into compositions. Furthermore, SUMMARY OF THE DISCLOSURE the disclosure provides microbial consortia, containing at least two members of the disclosed microorganisms, as well 0009. In some aspects, the present disclosure provides as methods of utilizing said consortia. Furthermore, the isolated microbes, including novel strains of microbes, pre disclosure provides for methods of modulating the rumen sented in Table 1 and/or Table 3. microbiome. 0010. In other aspects, the present disclosure provides isolated whole microbial cultures of the microbes identified STATEMENT REGARDING SEQUENCE in Table 1 and Table 3. These cultures may comprise LISTING microbes at various concentrations. 0011. In some aspects, the disclosure provides for utiliz 0003. The sequence listing associated with this applica ing one or more microbes selected from Table 1 and/or Table tion is provided in text format in lieu of a paper copy, and 3 to increase a phenotypic trait of interest in a ruminant. is hereby incorporated by reference into the specification. Furthermore, the disclosure provides for methods of modu The name of the text file containing the sequence listing is lating the rumen microbiome by utilizing one or more ASBI 002 02US SeqList ST25.txt. The text file is 893 kb, microbes selected from Table 1 and/or Table 3. was created on Oct. 31, 2016, and is being submitted 0012. In some embodiments, a microbial consortium electronically via EFS-Web. comprises at least two microbial strains selected from Table 1 and/or Table 3. In some embodiments, a microbial con BACKGROUND sortium comprises at least one microbial strain selected from Table 1 and/or Table 3. In a further embodiment, a microbial 0004. The global population is predicted to increase to consortium comprises at least two microbial strains, wherein over 9 billion people by the year 2050 with a concurrent each microbe comprise a 16S rRNA sequence encoded by a reduction in the quantity of land, water, and other natural sequence selected from SEQID NOs: 1-30 and 2045-2103 or resources available per capita. Projections indicate that the an ITS sequence selected from SEQ ID NOS:31-60 and average domestic income will also increase, with the pro 2104-2107. In an additional embodiment, a microbial con jected rise in the GDP of China and India. The desire for a sortium comprises at least one microbial strain, wherein diet richer in animal-source proteins rises in tandem with each microbe comprise a 16S rRNA sequence encoded by a increasing income, thus the global livestock sector will be sequence selected from SEQ ID NOs: 1-30 and 2045-2103, charged with the challenge of producing more milk using or an ITS sequence selected from SEQ ID NOS:31-60 and fewer resources. The Food and Agriculture Organization of 2104-2107. the United Nations predict that 70% more food will have to 0013. In some embodiments, the microbial consortia of be produced, yet the area of arable land available will the present disclosure comprise at least two microbial decrease. It is clear that the food output per unit of resource strains, wherein each microbe comprises a 16S rRNA input will have to increase considerably in order to Support sequence encoded by a sequence selected from SEQ ID the rise in population. NOs: 1-30, SEQ ID NOs:61-1988, or SEQ ID NOs:2045 0005 Milk and milk components from lactating rumi 2103; or an ITS sequences selected from SEQ ID NOS:31 nants are predominantly utilized in the preparation of food 60, SEQ ID NOs: 1989-2044, or SEQ ID NOS:2104-2107. stuffs in many different forms. Nevertheless, milk and milk 0014. In one embodiment, the microbial consortium com components find numerous alternative applications in non prises at least two microbial strains comprising Ascusb 7. food areas such as the manufacture of glues, textile fibers, Ascusb 32, Ascusf 45, and Ascusf 24. In a further embodi plastic materials, or in the production of ethanol or methane. ment, the microbial consortium comprises at least one There have been many strategies to improve milk production microbial Strain comprising Ascusb 7, Ascusb 32, Ascusf and content in ruminants through nutritional modulations, 45, and Ascusf 24.
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