Acetobacter Aceti and Kluyveromyces Lactis August 2016

Staff Assessment Report APP202920: to determine the new organism status of Acetobacter aceti and Kluyveromyces lactis August 2016

Purpose To determine if Acetobacter aceti and Kluyveromyces lactis are new organisms under Section 26 of the HSNO Act

Application number APP202920 Application type Statutory Determination Applicant Natural Solutions NZ Ltd Date formally received 21 July 2016 EPA Staff Assessment Report: Application APP202920

Executive Summary and Recommendation

Natural Solutions New Zealand Ltd has submitted the application APP202920 in order to seek a determination on the new organism status of Acetobacter aceti and Kluyveromyces lactis.

After reviewing the information provided by the applicant, EPA staff recommends the Hazardous Substances and New Organisms (HSNO) Decision-making Committee determine that Acetobacter aceti and Kluyveromyces lactis are not new organisms for the purpose of the HSNO Act.

EPA Staff Assessment Report: Application APP202920

Table of Contents

Executive Summary and Recommendation ...... 2

Table of Contents ...... 3

1. Introduction ...... 4

Purpose of this document ...... 4 The application ...... 4 2. Summary of information ...... 4

Acetobacter aceti ...... 4 Table 1: Summary of locations Acetobacter aceti has been isolated from: ...... 6 Kluyveromyces lactis ...... 6 Table 2: Summary of environments Kluyveromyces lactis has been isolated from: ...... 7 Ubiquity of microorganisms ...... 7 3. Evaluation against legislative criteria ...... 7

Acetobacter aceti ...... 8 Kluyveromyces lactis ...... 8 4. Comments from government agencies ...... 9

5. Overall findings and conclusion ...... 9

6. References ...... 10

Appendix 1: Correspondence ...... 13

Appendix 2: Comments from MPI ...... 14

Appendix 3: Decision path for section 26 determination ...... 16

Context ...... 16 Introduction ...... 16 Figure 1 Flowchart: Decision path for applications under Section 26 for determination as to whether an organism is a new organism ...... 17 Figure 1 Explanatory Notes ...... 18

EPA Staff Assessment Report: Application APP202920

1. Introduction

Purpose of this document

This document has been prepared by Environmental Protection Authority (EPA) staff to advise the Hazardous Substances and New Organisms (HSNO) Decision-making Committee (the Committee) of our assessment of application APP202920 submitted under the HSNO Act (the Act). This document discusses information provided in the application and various other sources.

The application

The application from Natural Solutions NZ Ltd was submitted under section 26 of the Act to determine whether Acetobacter aceti (Pasteur 1864) Beijerinck 1898 and Kluyveromyces lactis (Dombrowski) van der Walt 1971 are new organisms for the purpose of the Act. This application was formally received by the EPA on 27 July 2016.

The applicant had previously been importing these organisms contained within a kefir starter culture trading under the name Bravo manufactured in Switzerland by Silver Spring Sagl.

Prior to June 2016 the applicant had imported this product under the Import Health Standard for the Importation into New Zealand of Specified Animal Products and Biologicals (INEPROIC.ALL). This import health standard allows for the import of ‘commercially manufactured food cultures, enzymes or starters derived from or consisting of micro-organisms’ and ‘brewer’s yeast, baker’s yeast or any other yeast products in the food industry’ from any country. As of 27 August 2015 a clause was added to the import health standard stating that there may be additional requirements for microorganisms under the HSNO Act. Therefore the applicant contacted the EPA for a formal determination on Acetobacter aceti and Kluyveromyces lactis to be able to continue importing the starter culture product into New Zealand.

The applicant considers Acetobacter aceti and Kluyveromyces lactis to be naturally occurring in the New Zealand environment and widely used in the food industry. Therefore, the applicant considers these organisms to be not new for the purpose of the HSNO Act.

Our assessment considers the information contained within the application and any other relevant information that we found in scientific literature or elsewhere. 2. Summary of information

Acetobacter aceti

Acetobacter aceti bacteria are gram-negative, obligate aerobic rods which can oxidise ethanol into acetic acid (Matsushita et al 2005). Acetobacter aceti is ubiquitous in the environment, existing in alcoholic niches in flowers and fruit, and in soil and water. The main industrial use of A. aceti is for the production of vinegar (US EPA 1997).

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Worldwide, acetic acid bacteria (AAB) play a large part in the production and spoilage of wine. Acetobacter aceti is an AAB that is normally associated with grapes and wine. The exposure of wine to oxygen during the production process often leads to a proliferation of A. aceti bacteria. High numbers of A. aceti bacteria in wine results in a product with undesirable traits such as high volatile acidity and a vinegary taint (Du Toit et al 2005). Acetobacter aceti can be isolated from unspoiled grapes in low amounts, and from damaged grapes in much higher numbers (Guillamón and Mas, 2009). Work has also been done on the main species of AAB present on grapes/musts and wine. Acetobacter aceti is found throughout the production process (i.e. on grapes/must, mid fermentation and end fermentation) particularly on botrytized grapes1 in France, cabernet sauvignon in South Africa, and red Grenache in Spain (Unden and Frohlich, 2009). While we could find no evidence that A. aceti had been isolated in grapes and wine in New Zealand it is unlikely that New Zealand remains untouched by A. aceti given its ubiquity in the wine industry.

Commercially, A. aceti is commonly used to ferment alcohol into vinegar. There are several companies in New Zealand that produce vinegar for the domestic market. There are also companies that export vinegar to New Zealand. Dominion Yeast Company Ltd (DYC) was established in Christchurch in 1915 and produces a range of vinegar products. These products are made in New Zealand by natural fermentation using A. aceti (DYC, 2016). Another New Zealand based company, Ceres Organics, produces a raw apple cider vinegar that ferments New Zealand apples with A. aceti. This product is unpasteurised, unfiltered, and undiluted. Bragg Live Foods is an American company that produces a variety of organic foodstuffs. Braggs have been exporting raw apple cider vinegar to New Zealand since 2002. Braggs apple cider vinegar is unfiltered and unpasteurized, and is fermented with A. aceti (Appendix 1). These three companies, and other vinegar producers in New Zealand (Coraltree Organics, Artisan Vinegar, Chantal Organics, Goulter’s Vinegar Products) mention that, over time, their products may develop the ‘mother of vinegar’. The mother is a non-toxic film that accumulates on the surface of vinegar during oxidisation and is made up of AAB and cellulose. Acetobacter aceti, along with several other species from the Acetobacter, Gluconacetobacter and Komagataeibacter genera are commonly detected in the mother of vinegar (Yetiman and Kesmen 2015). All of these products are widely available in supermarkets across New Zealand.

Table 1 indicates that A. aceti is a cosmopolitan species, and has been identified in a range of environments around the world, many of which occur in New Zealand. Acetobacter aceti is commonly associated with vinegar and wine production, two activities which are carried out across New Zealand. This strongly indicates that A. aceti is a ubiquitous organism that should be considered present in New Zealand.

1 Botrytis cinerea is a fungal infection that causes grapes to dehydrate and shrivel into raisins. Wines made from these infected grapes are sweet in nature.

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Table 1: Summary of locations Acetobacter aceti has been isolated from: Acetobacter aceti

Environment Country Reference

Fermented tea Taiwan Liu et al 1996

Alcohol and vinegar Pakistan Zahoor et al 2006

Cacao beans Trinidad Ostovar and Keeney 1973

Kefir Iran Motaghi et al 1997

Vinegar Japan Ohmori et al 1980

Grape pomace Portugal Rozes et al 2000

Bananas Nigeria Samuel et al 2016

Kluyveromyces lactis

Kluyveromyces lactis is a unicellular, lactose assimilating, budding yeast (Schaffrath and Breunig 2000). Kluyveromyces lactis is an aerobic organism with ovoid cells and is considered a model organism for studies in genetics and physiology (Dias et al 2012). This is due to its ability to produce commercially relevant proteins in the food, feed and pharmaceutical industries (Spohner et al 2016).

Commercially, K. lactis is used to produce enzymes like lactase and chymosin. The ability of K. lactis to degrade lactose in milk products into galactose and glucose is particularly helpful for producing products for lactose intolerant consumers. Chymosin is a milk clotting enzyme that forms the active constituent of cheese rennet (van Ooyen et al 2006; van den Berg et al 1990). In studies of yeasts and moulds isolated from cheese, K. lactis is often excluded as it is used specifically as a ripening culture and it is impossible to determine if it is a natural contaminants or a natural part of the production process (Lavoie, 2012).

Kluyveromyces lactis has been repeatedly isolated from raw milk (Elgadi et al 2008; Sugai et al 2009; Quigley et al 2013) and is a key member of the microflora in ripening cheese. In cheese production, Kluyveromyces lactis decreases the pH of whey and allows lactic acid bacteria to proliferate (Rodicio and Heinisch 2013). Kluyveromyces lactis and K. marxianus are the two most common yeasts isolated from kefir starter cultures (Bourrie et al 2016).

The only direct evidence of the presence of K. lactis in New Zealand in the scientific literature is from one study carried out in 1966 in the Taranaki region. This study references the presence of Saccharomyces lactis, the basionym for K. lactis, and examined the yeast flora present in pasture that was spray-irrigated with dairy factory wastes. Saccharomyces lactis was isolated from soil irrigated with dairy waste in Stratford, New Zealand (Di Menna, 1966).

The preceding evidence coupled with the detection of K. lactis around the globe (Table 2) indicates that K. lactis is a ubiquitous organism that is often found and purposely used in the dairy industry.

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Table 2: Summary of environments Kluyveromyces lactis has been isolated from: Kluyveromyces lactis

Environment Country Reference

Winery equipment South Africa van der Walt and Nel 1963

Slime flux of an oak tree Russia Shimizu et al 1985

Cheese Spain Padilla et al 2012

Creamery USA Roberto et al 1990

Cheese Italy Andrighetto et al 2000

Sugar factory Trinidad Nagassar-Mohit 1988

Yoghurt Australia Suriyarachchi and Fleet 1981

Ubiquity of microorganisms

Current scientific thinking suggests that non-symbiotic or free-living prokaryotic microorganisms are ubiquitous in their distribution and are only limited by their specific habitat requirements. This is attributed to their small size, high abundance and ability to rapidly adapt via horizontal gene transfer. Bacterial cells can be readily transported by natural processes (wind, water, migratory animals) as well as being dispersed by human vectors that have allowed non-symbiotic bacteria to disperse globally (Ehlers and Lear, 2014).

Acetobacter aceti has been isolated internationally from fermented products such as tea, vinegar, wine, and kefir (Table 1). Kluyveromyces lactis is most commonly isolated from dairy products such as milk and cheese.

These products and commercial starter cultures to make some of these products have been imported into New Zealand for a significant period of time. Moreover, the specific habitats where the two organisms are known to grow globally have been present in the New Zealand environment prior to 29 July 1998 and those fermented and dairy-based products have been made in New Zealand since before this date.

The ubiquity of Acetobacter aceti and Kluyveromyces lactis in geographically distinct environments internationally supports the presence of these species in New Zealand prior to 29 July 1998. 3. Evaluation against legislative criteria

For an organism to be determined as “not new” under section 26 of the Act, the organism must be shown to lie outside the parameters of the definition of a new organism as defined in section 2A(1) of the Act:

A new organism is-

a) an organism belonging to a species that was not present in New Zealand immediately before 29 July 1998:

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b) an organism belonging to a species, subspecies, infrasubspecies, variety, strain, or cultivar prescribed as a risk species, where that organism was not present in New Zealand at the time of promulgation of the relevant regulation:

c) an organism for which a containment approval has been given under this Act:

ca) an organism for which a conditional release has been given: cb) a qualifying organism approved for release with controls:

d) a genetically modified organism:

e) an organism that belongs to a species, subspecies, infrasubspecies, variety, strain, or cultivar that has been eradicated from New Zealand.

The decision pathway for a section 26 determination is outlined in Appendix 3.

Acetobacter aceti

Section 2A(1)(a) of the Act states that a new organism must belong to “a species that was not present in New Zealand immediately before 29 July 1998”. We have evaluated the information regarding A. aceti against this criterion.

The following Act criteria were not applicable to this determination as the bacterium Acetobacter aceti under consideration in this application:  has not been prescribed as a risk species (section 2A(1)(b))  is not a genetically modified organism (section 2A(1)(d))  has not been eradicated from New Zealand (section 2A(1)(e)).

However, Acetobacter aceti is currently listed on the containment approval NOC99014. NOC99014 was approved in 2000 under the Transitional Provisions for micro-organisms lawfully in use (section 259 of the HSNO Act, expired 29 July 2001). This approval was intended to capture all the microorganisms in the International Collection of Microorganisms from Plants (IMCP) held by Landcare Research. The original application, in accordance with section 259 of the HSNO Act, sought to maintain in containment, and to re-import into containment, microorganisms held as reference specimens for national and international research programmes.

Subsequent evidence presented in this report indicates Acetobacter aceti is present in the New Zealand environment and should be removed from the containment approval NOC99014.

Kluyveromyces lactis

The following Act criteria were not applicable to this determination as the yeast Kluyveromyces lactis under consideration in this application:

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 has not been prescribed as a risk species (section 2A(1)(b))  has not been approved to be held in containment or released with controls (sections 2A(1)(c), (ca) and (cb))  is not a genetically modified organism (section 2A(1)(d))  has not been eradicated from New Zealand (section 2A(1)(e)). 4. Comments from government agencies

In accordance with section 58(1)(c) of the Act, and clauses 2(2)(e) and 5 of the Methodology, the Department of Conservation (DOC) and the Ministry for Primary Industries (MPI) were notified and provided with the opportunity to provide further information on the application.

DOC advised on the receipt of this notification that they had no information or evidence that would help to determine whether Acetobacter aceti and Kluyveromyces lactis were present in NZ prior to 29 July 1998.

MPI does not have any information indicating the presence or absence of Acetobacter aceti or Kluyveromyces lactis prior to 29 July 1998. MPI concur that A. aceti is ubiquitous in the environment and requested further evidence for both A. aceti and K. lactis to strengthen the case for presence (for full comments see Appendix 2). We believe these comments have been addressed in the summary of information that forms section two of this document. 5. Overall findings and conclusion

Based on the information available we consider that there is sufficient weight of evidence to conclude that Acetobacter aceti and Kluyveromyces lactis were present in New Zealand prior to 29 July 1998 and should be deemed as not new for the purpose of the Act.

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6. References

Andrighetto, C., Psomas, E., Tzanetakis, N., Suzzi, G., & Lombardi, A. (2000). Randomly amplified polymorphic DNA (RAPD) PCR for the identification of yeasts isolated from dairy products. Letters in applied microbiology, 30(1), 5-9.

Bourrie, B. C., Willing, B. P., & Cotter, P. D. (2016). The Microbiota and Health Promoting Characteristics of the Fermented Beverage Kefir. Frontiers in microbiology, 7.

Di Menna, M. E. (1966). Yeasts in soils spray-irrigated with dairy factory wastes. New Zealand Journal of Agricultural Research, 9(3), 576-589.

Dias, O., Gombert, A. K., Ferreira, E. C., & Rocha, I. (2012). Genome-wide metabolic (re-) annotation of Kluyveromyces lactis. BMC genomics, 13(1), 1.

Du Toit, W. J., Pretorius, I. S., & Lonvaud‐Funel, A. (2005). The effect of sulphur dioxide and oxygen on the viability and culturability of a strain of Acetobacter pasteurianus and a strain of Brettanomyces bruxellensis isolated from wine. Journal of Applied Microbiology, 98(4), 862-871.

DYC, 2016. Vinegar FAQs. Retrieved 12 August 2016. http://www.cleverliving.nz/faqs/

Ehlers C, Lear G 2014. The Biogeography of Environmental Microorganisms EPA Contract Reference Number: AAN2014-130.

Elgadi, Zeinab AM, Warda S. Abdel Gadir, and Hamid A. Dirar. "Isolation and identification of lactic acid bacteria and yeast from raw milk in Khartoum State (Sudan)." Research Journal of Microbiology 3.3 (2008): 163-168.

Guillamón, J. M., & Mas, A. (2009). Acetic acid bacteria. In Biology of Microorganisms on Grapes, in Must and in Wine (pp. 31-46). Springer Berlin Heidelberg.

Lavoie, K., Touchette, M., St-Gelais, D., & Labrie, S. (2012). Characterization of the fungal microflora in raw milk and specialty cheeses of the province of Quebec. Dairy science & technology, 92(5), 455-468.

Liu, C. H., Hsu, W. H., Lee, F. L., & Liao, C. C. (1996). The isolation and identification of microbes from a fermented tea beverage, Haipao, and their interactions during Haipao fermentation. Food Microbiology, 13(6), 407-415.

Matsushita, K., Inoue, T., Adachi, O., & Toyama, H. (2005). Acetobacter aceti possesses a proton motive force-dependent efflux system for acetic acid. Journal of bacteriology, 187(13), 4346-4352.

Motaghi, M., Mazaheri, M., Moazami, N., Farkhondeh, A., Fooladi, M. H., & Goltapeh, E. M. (1997). Kefir production in Iran. World Journal of Microbiology and Biotechnology, 13(5), 579-581.

Ohmori, S., Masai, H., Arima, K., & Beppu, T. (1980). Isolation and identification of acetic acid bacteria for submerged acetic acid fermentation at high temperature. Agricultural and Biological Chemistry, 44(12), 2901-2906.

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Ostovar, K., & Keeney, P. G. (1973). Isolation and characterization of microorganisms involved in the fermentation of Trinidad's cacao beans. Journal of Food Science, 38(4), 611-617.

Padilla, B., Ruiz-Matute, A. I., Belloch, C., Cardelle-Cobas, A., Corzo, N., & Manzanares, P. (2012). Evaluation of oligosaccharide synthesis from lactose and lactulose using β-galactosidases from Kluyveromyces isolated from artisanal cheeses. Journal of agricultural and food chemistry, 60(20), 5134-5141.

Quigley, L., O'Sullivan, O., Stanton, C., Beresford, T. P., Ross, R. P., Fitzgerald, G. F., & Cotter, P. D. (2013). The complex microbiota of raw milk. FEMS microbiology reviews, 37(5), 664-698.

Roberto, C. O., María, E. Z. O., Valentín, M. R., Gloria, A. J., de Alba Tuena, C., & Aurora, B. L. (1990). Restriction site variation, length polymorphism and changes in gene order in the mitochondrial DNA of the yeast Kluyveromyces lactics. Antonie van Leeuwenhoek, 58(4), 227-234.

Rodicio, R., & Heinisch, J. J. (2013). Yeast on the milky way: genetics, physiology and biotechnology of Kluyveromyces lactis. Yeast, 30(5), 165-177.

Rozès, N., Barón, A. M., Martínez, M. J. T., Miranda, C. R., Icart, M. P., Ruiz, A., ... & Beltrán, G. (2000). Seguimiento e identificación de los microorganismos presentes durante la elaboración de los vinos. Alimentación, equipos y tecnología, 19(2), 95-100.

Samuel, O., Lina, J., & Ifeanyi, O. (2016). Production of Vinegar from Oil-palm Wine Using Acetobacter aceti Isolated from Rotten Banana Fruits. Universal Journal of Biomedical Engineering, 4(1), 1-5.

Schaffrath, R., & Breunig, K. D. (2000). Genetics and molecular physiology of the yeast Kluyveromyces lactis. Fungal Genetics and Biology, 30(3), 173-190.

Shimizu, K., Adachi, T., Kitano, K., Shimazaki, T., Totsuka, A., Hara, S., & Dittrich, H. H. (1985). Killer properties of wine yeasts and characterization of killer wine yeasts. Journal of fermentation technology, 63(5), 421-429.

Spohner, S. C., Schaum, V., Quitmann, H., & Czermak, P. (2016). Kluyveromyces lactis: An emerging tool in biotechnology. Journal of biotechnology, 222, 104-116.

Sugai, M., Takakuwa, N., Ohnishi, M., Urashima, T., & Oda, Y. (2009). Characterization of sterol lipids in Kluyveromyces lactis strain M-16 accumulating a high amount of steryl glucoside. Journal of Oleo Science, 58(2), 91-96.

Suriyarachchi, V. R., & Fleet, G. H. (1981). Occurrence and growth of yeasts in yogurts. Applied and environmental microbiology, 42(4), 574-579.

Unden, G., & Fröhlich, J. (2009). Biology of Microorganisms on Grapes, in Must and in Wine (pp. 3-29). H. König (Ed.). Heidelberg: Springer.

US EPA, 1997. "Acetobacter aceti Final Risk Assessment. Retrieved 15 August 2016. http://widit.knu.ac.kr/epa/ebtpages/Pollutants/Toxics/Microorganisms/siteout/s1out2.htm

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van den Berg, J. A., van der Laken, K. J., van Ooyen, A. J., Renniers, T. C., Rietveld, K., Schaap, A., ... & Richman, M. (1990). Kluyveromyces as a host for heterologous gene expression: expression and secretion of prochymosin. Nature Biotechnology, 8(2), 135-139. van der Walt, J. P., & Nel, E. E. (1963). Saccharomyces vanudenii nov. spec. Mycopathologia, 20(1), 71-74. van Ooyen, A. J., Dekker, P., Huang, M., Olsthoorn, M. M., Jacobs, D. I., Colussi, P. A., & Taron, C. H. (2006). Heterologous protein production in the yeast Kluyveromyces lactis. FEMS yeast research, 6(3), 381-392.

Yetiman, A. E., & Kesmen, Z. (2015). Identification of acetic acid bacteria in traditionally produced vinegar and mother of vinegar by using different molecular techniques. International journal of food microbiology, 204, 9-16.

Zahoor, T., Siddique, F., & Farooq, U. (2006). Isolation and characterization of vinegar culture (Acetobacter aceti) from indigenous sources. British Food Journal, 108(6), 429-439.

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Appendix 1: Correspondence Dear Sarah Fish, Thank you for your e-mail. Yes, the bacteria is called acetic acid bacteria from the Acetobacter Aceti family. Bragg has been exporting ACV to New Zealand since 2002.

Blessings of health! Bragg Customer Service Tel: 1-(800)-446-1990

From: [email protected] [mailto:[email protected]] Sent: Thursday, August 11, 2016 2:46 PM To: [email protected] Subject: Web Contacts Request Bragg Product Info-q

Web Contact Request Bragg Product Info-q

------Name: Sarah Fish Email: [email protected] Phone: 6444745581 Subject: Bragg Product Info-q Address: Level 10 215 Lambton Quay Wellington 6011 New Zealand Comments: Hello, Are you able to confirm whether Braggs raw apple cider contains the bacteria 'Acetobacter aceti'? Are you also able to tell me how long you have been exporting apple cider to New Zealand? Kind Regards, Sarah ------If there are any problems with this form or its syntax please email [email protected]

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Appendix 2: Comments from MPI

Comments Form to the EPA for New Organism Applications

Application Code(s): APP202920 Applicant Name: Natural Solutions NZ Ltd Application Categories: Determination of whether an organism is a new organism Application Title(s): Determination of whether Acetobacter aceti and Kluyveromyces lactis are new organisms. EPA Applications Contact: Sarah Fish Date: 28 July 2016 MPI Response Coordinator: Barry Wards Option to Speak in support of these No comments: BASIS ON WHICH COMMENT IS PROVIDED

MPI submits these comments for consideration to the EPA on the following (where relevant to the type of application):  Clarity of information;  Information that MPI considers should be taken into consideration by the EPA;  Adequacy of the proposed containment system, including suggestions for controls and amendments to proposed controls; and  Enforceability of any proposed controls.

Matters relating to the application that are not within the scope of these comments may be provided to the EPA separately.

Comments

General . The applicant has applied to the EPA for a determination of whether the bacterial species Acetobacter aceti and the fungal species Kluyveromyces lactis are new organisms. A. aceti . MPI is unable to provide any information indicating that A. aceti was present in New Zealand prior to 29 July 1998. MPI databases are unable to be searched to determine if A. aceti has been specifically imported. Since the organism has been used extensively in the manufacture of vinegar for many years, it may well have been imported prior to 29 July 1998. We note that under the import health standard for the Importation into New Zealand of Specified Animal Products and Biologicals (INEPROIC.ALL) no permit to import for food starter cultures is required. However, we believe that none of this information is sufficient to conclude that A. aceti was imported into New Zealand prior to 29 July 1998. . MPI notes the following points regarding the application: - The applicant has stated that “A. aceti has been used extensively in NZ since 1915 for the commercial production of vinegar by the DYC Vinegar Company, as confirmed by the email of Bettesworth (7 June 2016)”. The email from C. Bettesworth does not actually state this – it only states that all DYC vinegars are naturally fermented with A.

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aceti. There is no further information as to how this achieved or for how long a particular process has been employed. We suggest the applicant asks the DYC Vinegar manufacturers (a) whether A. aceti has been used prior to 29 July 1998, (b) where the A. aceti been sourced from (ie, on fermentable goods or imported), (c) do manufactured DYC Vinegars produced prior to 29 July 1998 contain viable A. aceti, and (c) can it confirm that A. aceti has been imported prior to 29 July 1998. - MPI concurs that Bragg raw apple cider (being unfiltered, unheated and unpasteurized) is likely to contain viable A. aceti. While the applicant states that this product is imported and sold widely throughout NZ, they have not supplied any evidence to indicate that this occurred prior to 29 July 1998. MPI suggests that the applicant contact Braggs and request this information. . While MPI concurs that A. aceti is ubiquitous in the environment, this statement is based on limited overseas reports. We suggest that reliance on further information, as suggested above, would strengthen a case for presence in NZ prior to 29 July 1998. K. lactis . MPI is unable to provide any further information indicating that K. lactis was present in New Zealand prior to 29 July 1998 but makes the following points regarding the application: - While the applicants cite the K. lactis listing in the Landcare Research website as being present, indigenous and non-endemic, there is no information in that website to support this. - Similarly, while Pennycook & Galloway (Introduction to Fungi of New Zealand, 2004) cite K. lactis in the “Checklist of New Zealand Fungi”, they provide no information to support this – MPI suggests that the listing is based on the Landcare Research website listing. - The report of Di Menna (1966) identifies that Saccharomyces lactis Dobrowski (syn. K. lactis) was retrieved from soil in Stratford. Di Menna also notes that “species which had colonised the irrigant in the holding tanks and sprinkler systems were not able to persist in soil, nor upon leaves of pasture plants” and “no yeasts could be recovered from a sample of irrigant taken at the factory”. Irrespective of this, the K. lactis must have come from somewhere; therefore, MPI suggests that Di Menna provides evidence that K. lactis was present in New Zealand prior to 29 July 1998, regardless of where it was located or whether it persisted.

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Appendix 3: Decision path for section 26 determination

Context

This decision path describes the decision-making process for applications under Section 26 for determination as to whether an organism is a new organism.

Introduction

The purpose of the decision path is to provide the HSNO decision maker2 with guidance so that all relevant matters in the HSNO Act and the Methodology have been addressed. It does not attempt to direct the weighting that the HSNO decision maker may decide to make on individual aspects of an application.

In this document ‘section’ refers to sections of the HSNO Act, and ‘clause’ refers to clauses of the Methodology.

The decision path has two parts –

 Flowchart (a logic diagram showing the process prescribed in the HSNO Act and the Methodology to be followed in making a decision), and  Explanatory notes (discussion of each step of the process). Of necessity the words in the boxes in the flowchart are brief, and key words are used to summarise the activity required. The explanatory notes provide a comprehensive description of each of the numbered items in the flowchart, and describe the processes that should be followed to achieve the described outcome.

For proper interpretation of the decision path it is important to work through the flowchart in conjunction with the explanatory notes.

2 The HSNO decision maker refers to either the EPA Board or any committee or persons with delegated authority from the Board.

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Figure 1 Flowchart: Decision path for applications under Section 26 for determination as to whether an organism is a new organism

For proper interpretation of the decision path it is important to work through the flowchart in conjunction with the explanatory notes.

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Figure 1 Explanatory Notes

Item 1 Review the content of the application and all relevant information Review the application, Agency advice and any relevant information held by other Agencies, and advice from experts. Determine whether further information is required.

Item 2 Is this information sufficient to proceed? Review the information and determine whether or not there is sufficient information available to make a decision.

Item 3: Seek additional information If the HSNO decision maker considers that further information is required, then this may be sought either from the applicant (if there is an external applicant) or from other sources. If the HSNO decision maker considers that the information may not be complete but that no additional information is currently available, then the HSNO decision maker may proceed to make a determination3. If the application is not approved on the basis of lack of information (or if the organism is considered new) and further information becomes available at a later time, then the HSNO decision maker may choose to revisit this determination. In these circumstances the HSNO decision maker may choose to adopt a precautionary approach under section 7 of the Act.

Item 4: Identify scope of organism description The identification of the organism must be at an appropriate taxonomic classification. For applications involving potentially genetically modified organisms, the organism should be identified by describing the host organism and the processes to which it has been subjected to (for example injection with a non-replicative, non-integrative plasmid DNA vaccine).

Item 5:

Is it a GMO? Determine whether the organism is a GMO using the definitions in Section 2 of the Act and in the Hazardous Substances and New Organisms (Organisms Not Genetically Modified) Regulations 1998.

Item 6: Is the organism known to have been present in NZ immediately before 29 July 1998? Determine on the basis of the available information whether on balance of probabilities the organism is known to belong to a species that was present in New Zealand immediately prior to 29 July 1998. For the purposes of making a section 26 determination an organism is considered to be present in New Zealand if it can be established that the organism was permanently existing in New Zealand and was not present solely by way of being contained in a recognised safekeeping facility, immediately prior to 29 July 1998. The key phrases ‘permanently existing, ‘recognised

3 Alternatively the application may lapse for want of information.

EPA Staff Assessment Report: Application APP202920

safekeeping facility’ and ‘immediately’ are defined in the Protocol Interpretations and Explanations of Key Concepts

Item 7: Is it prescribed as a risk species? Determine whether the organism has been prescribed as a risk species by regulation established under section 140(1)(h) of the Act. Note: at this point it may become apparent that the organism is an unwanted organism under the Biosecurity Act. If this is the case, then MAF BNZ and DOC may be advised (they may already have been consulted under items 1, 2 and 3).

Item 8: Was it present when prescribed? If the organism is prescribed as a risk species, determine whether it was present when it was prescribed. The organism is a new organism if it was not present in New Zealand at the time of the promulgation of the relevant regulation (Section 2A (1)(b) of the Act).

Item 9: Is it known to have been previously eradicated? Determine whether the organism is known to have been previously eradicated. Eradication does not include extinction by natural means but is considered to be the result of a deliberate act (see the interpretation in the Protocol Interpretations and Explanations of Key Concepts1.

Item 10: Has HSNO release without conditions approval been given under section 38 or 38I of the Act? If a HSNO release approval has been given under section 35 of the Act, then the organism remains a new organism. If a release approval has been given under section 38 of the Act then the organism is not a new organism. If a release approval has been given under section 38I of the Act, then if the approval has been given with controls then the organism remains a new organism, however, if this approval has been given without controls then it is not a new organism.