Page 1 of 27 Heat-killed M. aurum Aogashima (Brand name: Au+) Novel food Ingredient Application By Solution Sciences Ltd. Page 2 of 27 1. Contents Table 1: Abbreviations ........................................................................................................................................ 3 Table 2: List of Annexes ...................................................................................................................................... 4 Table 3: List of References .................................................................................................................................. 5 1. Administrative Data .................................................................................................................................... 8 Notes on confidentiality .................................................................................................................................. 8 2. General Introduction and Description of Mycobacterium aurum Aogashima ............................................ 9 I The purpose of tHe novel food ingredient .............................................................................................. 9 3. Identification of essential information requirements ............................................................................... 11 4. Consultation of structured schemes ......................................................................................................... 12 I Structured ScHeme I: Specification of tHe Novel Food ingredient ........................................................ 12 i. Isolate origin, identity and pHysical properties ................................................................................. 12 ii. Isolate identification and cHaracterisation ........................................................................................ 13 iii. Genetic and microbiological identity of Mycobacterium aurum Aogashima .................................... 14 iv. Assessment of Mycobacterium aurum Aogashima isolate identity, cHaracterisation and microbial safety by microbiological techniques ........................................................................................................ 15 v. Specification and composition of Heat-killed M. aurum Aogashima ingredient ............................... 17 II Structured ScHeme II: Effect of tHe production process applied to tHe novel food ingredient. ........... 18 III Structured ScHeme III: History of tHe organism used as tHe source of tHe novel food ingredient ....... 20 IV– VIII Structured ScHemes: Not relevant to Heat killed M. aurum Aogashima (non-GMM) ...................... 22 IX Structured ScHeme IX: Anticipated intake/extent of use of tHe novel food ingredient ........................ 22 X Structured ScHeme X: Information from previous Human exposure to tHe novel food ingredient or its source ............................................................................................................................................................ 24 XI Structured ScHeme XI: Nutritional information on tHe novel food ingredient ..................................... 24 XII Structured ScHeme XII: Microbiological information on tHe novel food ingredient ............................. 24 XIII Structured ScHeme XIII: Toxicological information on the novel food ingredient ................................ 24 5. Evaluations and conclusion by applicant .................................................................................................. 27 Page 3 of 27 Table 1: Abbreviations ACNFP Advisory Committee on Novel Foods and Processes M. aurum Mycobacterium aurum ATCC American Type Culture Collection CFU Colony-Forming Units DNSIYC Diet and Nutrition Survey of Infants and Young CHildren EC European Community EFSA European Food Safety Authority EU European Union FAO Food and Agriculture Organisation GLP Good Laboratory Practice GMM Genetically Modified Microorganism GMO Genetically Modified Organism GMP Good Manufacturing Practice GVP Good PHarmacovigilance Practice HACCP Hazard Analysis & Critical Control Points HSE Health and Safety Executive NCTC National Collection of Type Cultures (UK) NDNS National Diet and Nutrition Survey NDNS1-3 National Diet and Nutrition Survey - Years 1-3 NF Novel Food NOAEL No Observed Adverse Effect Level NOEL No Observed Effect Level OECD Organisation for Economic Co-operation and Development OTC Over THe Counter PAMP Pathogen-Associated Molecular Pattern PCR Polymerase CHain Reaction QC Quality Control QPSNCTC Qualified Presumption of Safety National Collection of Type Cultures (UK) RASFFATCC Rapid Alert System For Food American Type Culture Collection TAMC Total Aerobic Microbial Counts TOC Total Organic Carbon TYMC Total Yeast & Mould Counts WGS Whole Genome Sequencing WHO World Health Organisation Page 4 of 27 Table 2: List of Annexes Annex Authors Year Full Title Study Code/Site FurtHer Details Annex 1 BioElpida / 2014 RNA16s tests results BEA-M.aurum-P14- Strain identification Bactup 14-04_27F CONFIDENTIAL Annex 2 Dr. L Rosa 2015 Mycobacterial genome Report Aurum CH- Strain identification Brunet assembly and R&D-01 CONFIDENTIAL Dr. D Simpson comparison: Mycobacterium aurum NCTC10437/ATCC2336 6 vs Mycobacterium aurum Aogashima Annex 3 Aurum 2015 Mycobacteria in water Aurum Switzerland Review of the natural Switzerland environments of mycobacteria Annex 4 BioElpida 2014 Manufacturing and BioElpida Manufacturing&QC quality control processes CONFIDENTIAL processes for Au+ Annex 5 BioElpida 2015 Certificates of analysis BioElpida Certificates of analysis of Au+ of 3 batches of Au+ CONFIDENTIAL Annex 6 Sequani 2015 14-Day oral dose range BIT0001 Toxicological data on finding test Heat killed M. aurum CONFIDENTIAL Aogashima Annex 7 Sequani 2015 SubcHronic 90-day oral BIT0002 Toxicological data on toxicity study Heat killed M. aurum CONFIDENTIAL Aogashima Annex 8 Sequani 2015 Bacterial reverse BIT0003 Toxicological data on mutation test Heat killed M. aurum CONFIDENTIAL Aogashima Annex 9 Covance 2015 Human LympHocyte BIT0004 Toxicological data on Micronucleus test Heat killed M. aurum CONFIDENTIAL Aogashima Annex 10 Aurum 2015 Hypersensitivity Aurum Switzerland Review of evidence Switzerland reactions induced by that mycobacteria environmental encountered in tHe mycobacteria natural environment do not cause hypersensitivity reactions Annex 11 References All scientific literature used in the dossier CONFIDENTIAL Page 5 of 27 Table 3: List of References Reference Authors Year Full Title Study Code/Site FurtHer Details Allergome Allergome allergome.org, http://www.allergo database database consulted on June me.org 2015 Brown et al. Brown K, 2012 Diet-induced dysbiosis Nutrient. 2012, 4, 2012 DeCoffe D, of the intestinal 1095-1119 Molcan E, microbiota and tHe Gibson DL. effects on immunity and disease Cambier et al. Cambier CJ, 2014 Host evasion and Cell. 2014 Dec 2014 Falkow S, exploitation scHemes 18;159(7):1497-509 Ramakrishnan L. of Mycobacterium tuberculosis. Dir. 2003/89 EU Commission 2003 Directive 2003/89/EC of the European Parliament & of tHe Council as regards indication of tHe ingredients present in foodstuffs Gey van 2006 Evolution and BMC Evol Biol. 2006 Gey van Pittius Pittius et al. expansion of tHe Nov 15;6:95. NC, Sampson 2006 Mycobacterium SL, Lee H, et al. tuberculosis PE and PPE multigene families and tHeir association witH tHe duplication of the ESAT-6 (esx) gene cluster regions. Gomez-SmitH 2015 Sulfate Reducing Environ Sci TecHnol. Gomez-SmitH et al., 2015 Bacteria and 2015 Jul CK., LaPara TM., Mycobacteria 21;49(14):8432-40 Hozalski RM, Dominate tHe Biofilm Communities in a Chloraminated Drinking Water Distribution System. Grange 2014 2014 Mycobacterium In Davies PDO, Grange JM. tuberculosis – the Gordon S, Davies G. organism. eds. Clinical Tuberculosis, 5tH Edition. London: CRC Press. Chp. 3, pp. 39-53. 2014 HSE Advisory 2013 The Approved List of Classifies Committee n Biological Agents Mycobacterium aurum Dangerous as Hazard Group 1 – PatHogens unlikely to cause human disease Page 6 of 27 Reference Authors Year Full Title Study Code/Site FurtHer Details KisHi 2011 KisHi Y. 2011 Chemistry of Proc Natl Acad Sci mycolactones, tHe U S A. 2011 Apr causative toxins of 26;108(17):6703-8. Buruli ulcer. Rahman et al. 2014 MBio. 2014 Nov Rahman SA, Comparative analyses 2014 4;5(6):e02020. SingH Y, KoHli S, of nonpathogenic, et al. opportunistic, and totally pathogenic mycobacteria reveal genomic and biochemical variabilities and highlight the survival attributes of Mycobacterium tuberculosis. Rook 2009 Rook GAW. 2009 Review series on Immunology Definition of helminths, immune 2009;126(1):3-11. pseudocommensal. modulation and tHe hygiene hypothesis: The broader implications of tHe hygiene hypothesis. Salminen and Saliminen S and 1992 Lactic acid bacteria in Digestive Diseases DeigHton, DeigHton M the gut in normal and 1992, 10, 227-238 1992 disordered states Simmon et al. Simmon KE, Low 2009 Phylogenetic Int J Syst Evol Phylogenetic analysis 2009 YY, Brown- analysis of Microbiol. of the type strain of Elliott BA, Mycobacterium aurum 2009;59:1371-5. Mycobacterium aurum Wallace RJ Jr, and Mycobacterium and identity of otHer Petti CA. neoaurum witH strains labeled redescription of Mycobacterium aurum M. aurum culture collection strains. Skerman et al. Skerman VBD, 1908 Approved lists of Int J Syst bacteriol The internationally 1980 McGowan V, bacterial names. 1980; 30: 225-420. approved bacterial SneatH PHA. names