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 waiver under section 59 of the HSNO Act
I request for the Authority to waive any legislative information requirements (i.e. concerning ☐ the information that has been supplied in my application) that my application does not meet (tick if applicable).
Please list below which section(s) of this form are relevant to the information waiver request:
December 2013 EPA0327 10
Application Form To obtain a determination of whether an organism is a new organism
Appendices and referenced material (if any) and glossary (if required)
REFERENCES
Bettesworth, C. (2016). DYC vinegar. Auckland: Wilson Consumer Products Limited.
Boekhout, T., & Kurtzman, C. P. (1996). Principles and Methods Used in Yeast Classification, and an Overview of Currently Accepted Yeast Genera. In K. Wolf, Nonconventional Yeasts in Biotechnology: A Handbook (pp. 42-43). Berlin Heidelberg: Springer.
Brannt, W. T. (1890). Theory of the Formation of Vinegar. In A Practical Treatise on the Manufacture of Vinegar and Acetates, Cider, and Fruit-wines (pp. 25-26). Philadelphia: https://books.google.co.nz/books?id=oDBDAAAAIAAJ.
Di Menna, M. E. (1966). Yeasts in soils spray-irrigated with dairy factory wastes. New Zealand Journal of Agriculture Research, 576-589.
Landcare Research. (2016, June 29). Kluyveromyces lactis (Boidin, Abadie, J.L. Jacob & Pignal) Van der Walt, 1971. Retrieved from New Zealand Organisms Register: http://www.nzor.org.nz/names/9b0297a5- a5fd-47e1-999c-8a9645347598
Pennycook, S. R., & Galloway, D. J. (2004). Checklist of New Zealand "Fungi". In E. H. McKenzie, Introduction to Fungi of New Zealand (p. 444 & 448). Hong Kong: Fungal Diversity Press.
Rodicio, R., & Heinisch, J. J. (2013). Yeast on the milky way: genetics, physiology and biotechnology of Kluyveromyces lactis. Yeast, 165-177.
Sievers, M., & Swings, J. (2005). Genus | Acetobacter. In B. M. Trust., Bergey's Manual of Systematic Bacteriology, Second Edition, Volume Two, The Proteobacteria, Part C. (pp. 51-52). New York: Springer.
US EPA, O. o. (1997, March 7). US EPA, FINAL DECISION DOCUMENT: TSCA SECTION 5(H)(4) EXEMPTION FOR ACETOBACTER ACETI. Retrieved from United States Environmental Protection Agency: https://www.epa.gov/sites/production/files/2015-09/documents/fd001_0.pdf
Wesolowski-Louvel, M., Breunig, K. D., & Fukuhara, H. (1996). Kluyveromyces lactis. In K. Wolf, Nonconventional Yeasts in Biotechnology: A Handbook (p. 140). Berlin Heidelberg: Springer.
APPENDIX
APPENDIX-A_Bettesworth-7Jun2016-DYC-vinegar.pdf
APPENDIX-B_Sievers-2005-Acetobacter-pg51-52.pdf
APPENDIX-C_US-EPA-exemption-for-Acetobacter-aceti-1997.pdf APPENDIX-D_Brannt-1890-Theory-of-the-Formation-of-Vinegar-pg25-26.pdf APPENDIX-E_Rodicio-2013-Yeast-on-the-milky-way-pg165-177.pdf APPENDIX-F_Di-Menna-1966-Yeasts-in-soils-pg576-589.pdf APPENDIX-G_Wesolowski-Louvel-1996-Kluyveromyces-lactis-Page139-201.pdf APPENDIX-H_Boekhout-1996-Principles-and-Methods-Used-in-Yeast-Page42-43.pdf APPENDIX-I_McKenzie-2004-Fungi-of-New-Zealand-pg444+448.pdf
December 2013 EPA0327