APPLICATION FORM Section 26 Determination To obtain a determination of whether an organism is a new organism Send to Environmental Protection Authority preferably by email ([email protected]) or alternatively by post (Private Bag 63002, Wellington 6140) Payment must accompany final application; see our fees and charges schedule for details. Application Number APP202920 Date 30 June 2016 www.epa.govt.nz 2 Application Form To obtain a determination of whether an organism is a new organism 1. Applicant details 1.1. Applicant Company Name: (if applicable) Natural Solutions NZ Ltd Contact Name: Michael Kelly Job Title: Director Physical Address: 40 St Benedicts Street, Newton, Auckland 1010 Postal Address (provide only if not the same as the physical): Phone (office and/or mobile): +64-9-3681909 and +64-27-4141030 Fax: +64-28-2555-2601 Email: [email protected] 1.2. New Zealand agent or consultant (if applicable) Company Name: Contact Name: Job Title: Physical Address: Postal Address (provide only if not the same as the physical): Phone (office and/or mobile): Fax: Email: December 2013 EPA0327 3 Application Form To obtain a determination of whether an organism is a new organism 2. Information about the organisms 2.1. Name of organisms Identify the organisms as fully as possible Organism name: Acetobacter aceti Domain: Bacteria Phylum: Proteobacteria Class: Alphaproteobacteria Order: Rhodospirillales Family: Acetobacteraceae Genus: Acetobacter Species: Acetobacter aceti (Pasteur 1864) Beijerinck 1898 Synonyms: Mycoderma aceti Pasteur 1864, 125 Acetobacter aceti subsp. aceti (Pasteur 1864) De Ley and Frateur 1974. Cells ellipsoidal to rod shaped, straight or slightly curved, 0.6–0.9 x 1.0–4.0 lm, occurring singly, in pairs, or in chains. Motile by peritrichous flagella. Endospores are not formed. Gram negative. Obligately aerobic; metabolism is strictly respiratory with oxygen as the terminal electron acceptor. Never fermentative. For optimal growth; the pH is 4.0- 6.0 and the temperature is 28-30 °C. Oxidizes ethanol to acetic acid. Acetate is oxidized to CO2 and H2O (Sievers & Swings, 2005). December 2013 EPA0327 4 Application Form To obtain a determination of whether an organism is a new organism Organism name: Kluyveromyces lactis Kingdom: Fungi Phylum: Ascomycota Subphylum: Saccharomycotina Class: Saccharomycetes Order: Saccharomycetales Family: Saccharomycetaceae [as 'Saccharomycetes'] Genus: Kluyveromyces Species: Kluyveromyces lactis (Boidin, Abadie, J.L. Jacob & Pignal) Van der Walt, 1971 Synonyms: Candida sphaerica (B.W. Hammer & Cordes) S.A. Mey. & Yarrow, 1978 Guilliermondella lactis Boidin, Abadie, J.L. Jacob & Pignal, 1962 Kluyveromyces marxianus var. lactis Saccharomyces lactis Dombrowski, 1910 [as 'lactis α'] Torula sphaerica B.W. Hammer & Cordes, 1920 (Landcare Research, 2016) Kluyveromyces lactis has been isolated from milk and constitutes the predominant eukaryote during cheese production, which decreases the whey pH and thus paves the way for the lactic acid bacteria to thrive. It is a unicellular eukaryote, which divides by polar budding (i.e. it is an ascomyceteous yeast) and looks very much like a smaller version of ’the yeast’, Saccharomyces cerevisiae. In fact, it also prefers glucose as a carbon source, has a life cycle supported by two mating types, a stable haploid and a semistable diploid phase, and can be induced to produce tetrads by nutrient starvation (Rodicio & Heinisch, 2013). Kluyveromyces lactis cells are spherical to oval, essentially aerobic, and usually grown at a temperature of 25-28°C. The upper limit is approximately 40°C (Wesolowski-Louvel, Breunig, & Fukuhara, 1996). December 2013 EPA0327 5 Application Form To obtain a determination of whether an organism is a new organism 2.2. Regulatory status of the organisms Is any of the organisms that is the subject of this application also the subject of: An innovative medicine application as defined in section 23A of the Medicines Act 1981? ☐ Yes ☒ No An innovative agricultural compound application as defined in Part 6 of the Agricultural Compounds and Veterinary Medicines Act 1997? ☐ Yes ☒ No December 2013 EPA0327 6 Application Form To obtain a determination of whether an organism is a new organism 3. Evidence regarding whether the organism meets the definition of a new organism Acetobacter aceti Acetobacter aceti is a benign microorganism that is ubiquitous in the environment, existing in soil, water, flowers, fruits, honey bees, vinegar and kefir. It has a long history of safe use in the fermentation industry for the production of acetic acid from alcohol. It is not pathogenic to humans or animals. Although it often comes in contact with humans due to its widespread presence in the environment, it does not colonize human skin nor does it inhabit the human body. There are no reports in the literature suggesting any allergic or immunologic responses to the bacterium that has been used for decades in fermentation facilities. Releases of this microorganism to the environment through fermentation uses would not pose any significant ecological hazards, because this microorganism is ubiquitous in the environment and it is not pathogenic to animals or plants. (US EPA, 1997) Acetobacter aceti has been used extensively in New Zealand (since 1915) for the commercial production of vinegar by the DYC Vinegar Company as confirmed by their technical manager Claudia Bettesworth in an email on the 7th of June 2016 (Bettesworth, 2016). Bragg raw apple cider vinegar, which is imported and sold widely throughout New Zealand, is unfiltered, unheated and unpasteurized. Bragg documents on their website and on their product label, that this product contains the “mother of vinegar”. This mother of vinegar is used for the fabrication of vinegar on a large scale and the organisms to which the term mother of vinegar has been applied, were named by Louis Pasteur to be Mycoderma aceti (Brannt, 1890). Mycoderma aceti is Acetobacter aceti (Sievers & Swings, 2005). In summary Acetobacter aceti is a benign microorganism that is ubiquitous in the environment and further established in New Zealand by the DYC Vinegar Company and Bragg raw apple cider vinegar. It has value in the New Zealand economy for the large scale production of vinegar. December 2013 EPA0327 7 Application Form To obtain a determination of whether an organism is a new organism Kluyveromyces lactis Species of Kluyveromyces are isolated from soil, water, fruit and other plant materials, tree fluxes, dairy products, drosophila and occasionally from clinical specimens (Boekhout & Kurtzman, 1996). The natural habitat of Kluyveromyces lactis is diverse, but many strains were originally isolated from milk-derived products in which the major carbon source is lactose (Wesolowski-Louvel, Breunig, & Fukuhara, 1996). According to the Landcare Research website the biostatus of Kluyveromyces lactis in New Zealand is Present, Indigenous and Non-endemic (Landcare Research, 2016). Margaret E. Di Menna isolated Kluyveromyces lactis as Saccharomyces lactis Dombrowski in the soil from Stratford in Taranaki in 1966 (Di Menna, 1966). Kluyveromyces lactis and Candida sphaerica are listed in the “Checklist of New Zealand Fungi” in the authoritative “Introduction to Fungi of New Zealand” edited by Eric H.C. McKenzie of Landcare Research (Pennycook & Galloway, 2004). Due to its traditional employment in cheese production, Kluyveromyces lactis has a GRAS (generally regarded as safe) status and thus is also an attractive production organism in the food industry. It is also the natural choice for the production of ß-galactosidase, which is used to remove lactose from milk for the growing number of consumers with lactose intolerance. Moreover, the enzyme is applied in the production of cheese and yoghurt and by the ice-cream industry (Rodicio & Heinisch, 2013). In summary Kluyveromyces lactis is a benign microorganism that is ubiquitous in the environment and of benefit to economies. December 2013 EPA0327 8 Application Form To obtain a determination of whether an organism is a new organism 4. Checklist This checklist is to be completed by the applicant Application Comments/justifications All sections of the application form completed ☒ Yes ☐ No or you have requested an information waiver (If No, please discuss with an under section 59 of the HSNO Act Advisor to enable your application to be further processed) Confidential data as part of a separate, ☐ Yes ☒ No identified appendix Supplementary optional information attached: Copies of additional references ☒ Yes ☐ No Relevant correspondence ☒ Yes ☐ No Administration Are you an approved EPA customer? ☐ Yes ☒ No If Yes are you an: Applicant: ☐ Agent: ☐ If you are not an approved customer, payment of fee will be by: Direct credit made to the EPA bank ☒ Yes ☐ No account (preferred method of payment) ☒ Payment to follow Date of direct credit: Cheque for application fee enclosed ☐ Yes ☒ No ☐ Payment to follow Electronic, signed copy of application e- ☒ Yes mailed to the EPA December 2013 EPA0327 9 Application Form To obtain a determination of whether an organism is a new organism Signature of applicant or person authorised to sign on behalf of applicant ☒ I am making this application, or am authorised to sign on behalf of the applicant or applicant organisation. ☒ I have completed this application to the best of my ability and, as far as I am aware, the information I have provided in this application form is correct. Signature Date 30th June 2016 Request for information