Staff Assessment Report

APP203045: To determine the new organism status of Lactobacillus sanfranciscensis, Lactobacillus rossiae, Lactobacillus crustorum, Lactobacillus curvatus, Lactobacillus pontis, Lactobacillus hilgardii, Lactobacillus paralimentarius,

Leuconostoc lactis, Pediococcus parvulus and Pediococcus acidilactici 19 April 2017

Purpose To determine if Lactobacillus sanfranciscensis, Lactobacillus rossiae, Lactobacillus crustorum, Lactobacillus curvatus, Lactobacillus pontis, Lactobacillus hilgardii, Lactobacillus paralimentarius, Leuconostoc lactis, Pediococcus parvulus and Pediococcus acidilactici are new organisms under Section 26 of the HSNO Act

Application number APP203045 Application type Statutory Determination Applicant The New Zealand Institute for Plant & Food Research Date formally received 22 November 2016 EPA Staff Assessment Report: Application APP203045

Executive Summary and Recommendation

The New Zealand Institute for Plant & Food Research has submitted an application in order to seek a determination on the new organism status of Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici.

After reviewing the information provided by the applicant, we recommend that the Hazardous Substances and New Organisms (HSNO) Decision-making Committee determines that Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus acidilactici and P. parvulus are not new organisms for the purpose of the HSNO Act.

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EPA Staff Assessment Report: Application APP203045

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 ...... 5

Ubiquity of microorganisms ...... 9 3. Evaluation against legislative criteria ...... 10

4. Comments from government agencies ...... 10

5. Overall findings and conclusion ...... 11

6. References ...... 12

Appendix 1: Decision path for section 26 determination ...... 14

Context ...... 14 Introduction ...... 14 Figure 1 Flowchart: Decision path for applications under Section 26 for determination as to whether an organism is a new organism ...... 15 Figure 1 Explanatory Notes ...... 16

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EPA Staff Assessment Report: Application APP203045

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 APP203045 submitted under the HSNO Act (the Act). This document discusses information provided in the application and other sources.

The decision path for this application can be found in Appendix One of this memo.

The application

The application, from New Zealand Institute for Plant & Food Research, was submitted under section 26 of the Act to determine whether the organisms listed below are new organisms for the purpose of the Act: - Lactobacillus sanfranciscensis (Weiss & Schillinger 1984)

- Lactobacillus rossiae (Corsetti et al. 2005)

- Lactobacillus crustorum (Scheirlinck et al. 2007)

- Lactobacillus curvatus (Abo-Elnaga and Kandler 1965)

- Lactobacillus pontis (Vogel et al. 1994)

- Lactobacillus hilgardii (Douglas et al. 1936)

- Lactobacillus paralimentarius (Cai et al. 1999)

- Leuconostoc lactis (Garvie 1960)

- Pediococcus parvulus (Gunther & White 1961)

- Pediococcus acidilactici (Lindner 1887)

This application was formally received by the EPA on 22 November 2016.

The applicant considers that all these species are naturally occurring in the New Zealand environment and were widely used in the food industry prior to the definition of what a new organism is in the HSNO Act. Therefore, the applicant submits that these organisms to be not new.

None of these bacteria represent a risk to New Zealand’s environment and are considered to have probiotic1 activity.

Our assessment includes information contained within the application and other relevant information found in scientific literature or elsewhere.

1 Probiotics are microorganisms that are believed to provide health benefits when consumed. They are usually found in dairy foods or dietary supplements containing live bacteria that replace or add to the beneficial bacteria normally present in the gastrointestinal tract.

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2. Summary of information

Taxonomy of Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici:

Domain: Bacteria

Phylum: Firmicutes

Class: Bacilli

Order: Lactobacillales Family: Lactobacillaceae and Leuconostocaceae

Genus: Lactobacillus or Pediococcus and Leuconostoc

The taxonomic order Lactobacillales belongs to the large group of lactic acid bacteria: Gram-positive organisms that produce lactic acid as a by-product of glucose metabolism through fermentation.

There are two groups of lactic acid bacteria, based on differences in their ability to ferment sugars: homofermentative species convert sugars mostly into lactic acid, and heterofermentative species

convert sugars into lactic acid, acetic acid, ethanol and carbon dioxide (Giraffa et al. 2010).

Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici can be found in a variety of ecological niches. A recent publication on the history of Lactobacillus species as probiotics states: “They are known constituents of the human gut and also occur widely in dairy, meat, plant and fermented products of commercial value. As a result of their ancient anthropological use in food preservation and their ability to rapidly ferment carbohydrates to lactic acid, they have become industrially important bacteria and are used in a myriad of food and agricultural fermentations worldwide. Their growth causes acidification of food material, preserving the product and imparting unique textures and flavours” (Bull et al. 2013).

The ability to colonize such a variety of habitats is a consequence of the metabolic versatility of this group of lactic acid bacteria. Hence, it is not unexpected that Lactobacillales bacteria have been used for decades in food preparation (Giraffa et al. 2010).

These bacteria are found in different environments and locations around the world (De Vuyst et al. 2014), and numerous publications attest to their global ubiquity. Table 1 lists, not exhaustively, the types of samples and the locations where each bacterium has been previously isolated.

The use of sourdoughs as natural starters in bread making can be traced to the second millennium B.C. (Cappelle et al. 2013). A sourdough starter is created by the fermentation of a mixture of flour (wheat, rye or maize) and water. Almost all the bacteria involved in this process are Lactobacillales (De Vuyst et al. 2014), including Lactobacillus sanfranciscensis, L. rossiae, L. crustorum,

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L. curvatus, L. pontis, L. paralimentarius, Leuconostoc lactis and Pediococcus acidilactici (Table 1 and references within).

Lactic acid bacteria are also used around the world to develop aroma and flavour in various dairy products (Vedamuthu 1994). Other examples include cheese making in Azerbaijan using Lactobacillus curvatus (Ahmadova et al. 2013), and in fermented mare milks and products derived from the milks in Mongolia (Yu et al. 2011) and in Italy (Giannino et al. 2009) using Leuconostoc lactis.

Lactobacillales are present in alcoholic and non-alcoholic beverages, for example, wine in Spain and Australia (Pediococcus parvulus) (Davis et al. 1986; Pérez-Martín et al. 2015), as well as South Africa (Lactobacillus hilgardii) (Du Plessis et al. 2004). Lactic acid bacteria are also present in rice wine in China (L. hilgardii) (Liu et al. 2016) and Korea (L. crustorum) (Kim et al. 2012), as well as in a traditional fermented beverage called boza in Turkey (Leuconostoc lactis and P. parvulus) (Heperkan et al. 2014).

There has been international trade in sourdough, dairy products, beverages, cereals and other fermented products since the early European settlements (Wirtz 2003). Furthermore, the presence of Lactobacillales species has been confirmed in many other studies (see Table 1).

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EPA Staff Assessment Report: Application APP203045 Table 1: Summary of samples and locations where bacterium strains have been isolated. Dates of Bacteria name Synonym Environment Country References records

Lactobacillus Lactobacillus 1971 Sourdough, rye, USA, Germany, (Sugihara et al. 1971; Kitahara et al. sanfranciscensis sanfrancisco 2005 wheat China, Italy, France 2005; De Vuyst et al. 2014) (Weiss and Schillinger 1984) (Kline and Sugihara 2014 1971)

2014 Lactobacillus rossiae Spelt, sourdough, Belgium, Italy, (De Vuyst et al. 2014; Elizaquível et al. Lactobacillus rossii 2016 chichi (maize) China 2015; Liu et al. 2016) ( Corsetti et al. 2005) 2015

Wheat/rye, 2007 (Scheirlinck et al. 2007; Kim et al. 2012; sourdough, dairy Lactobacillus crustorum 2016 Belgium, China, Sharafi et al. 2015; Gobbetti et al. 2016; None products, 2012 Iran, Korea, Estonia Liu et al. 2016) (Scheirlinck et al. 2007) makgeolli (rice 2015 alcohol) 2016 Italy, Belgium, (Lee et al. 2005; Ahmadova et al. 2013; Sourdough, Bacterium curvatum 2014 France, Japan, Sakai et al. 2013; De Vuyst et al. 2014; Lactobacillus curvatus kimchi, wheat, rye, 2005 Tunisia, Nigeria, Gobbetti et al. 2016; Mamhoud et al. (Troili-Petersson takanazuke, (Troili-Petersson 1903) 2013 Finland, China, 2016) 1903) cheese 2010 Portugal, Azerbaijan France, Italy, Wheat, rye, spelt, 2016 Lactobacillus pontis Germany, Denmark, (Muller et al. 2000; Viiard et al. 2013; None sorghum, 2013 Sudan, Botswana, Gobbetti et al. 2016) (Vogel et al. 1994) sourdough 2000 Estonia

2016 Lactobacillus hilgardii Lactobacillus Type II Wheat, kefir France, Germany, (Arena et al. 2013; Zanirati et al. 2015; 2015 ( Douglas and Cruess 1936) (Fornachon 1943) grains, wine Brazil, Argentina Gobbetti et al. 2016) 2013

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EPA Staff Assessment Report: Application APP203045 Table 1: Summary of samples and locations where bacterium strains have been isolated. Dates of Bacteria name Synonym Environment Country References records Lactobacillus kimchii Italy, France, (Yoon et al. 2000), 2016 Lactobacillus paralimentarius Wheat, rye, spelt, Greece, USA, (Yoon et al. 2000; Kitahara et al. 2005; Lactobacillus bobalius 2005 sourdough Belgium, China, Gobbetti et al. 2016; Liu et al. 2016) ( Cai et al. 1999) 2000 (Mañes-Lázaro et al. New Zealand* 2008)

Leuconostoc Sourdough, milk 2015 Leuconostoc lactis Italy, Mongolia, (Giannino et al. 2009; Yu et al. 2011; argentinum and dairy products, 2011 ( Garvie 1960) Argentina Elizaquível et al. 2015) (Dicks et al. 1993) maize (chicha) 2009

2009 2014 (Davis et al. 1986; Narváez-Zapata et al. Pediococcus parvulus Agave, boza, wine, Australia, Bulgaria, None 2010 2010; Heperkan et al. 2014; Osimani et (Gunther and White 1961) cabbage Japan, Mexico, 2015 Spain, Turkey al. 2015) 1986 Pediococcus lolii Sourdough, Chinese rice wine, (Pepe et al. 2013; Li et al. 2016; Lv et al. Pediococcus acidilactici (Doi et al. 2009) 2013 Italy, Tunisia, barley and triticale, 2016; Mamhoud et al. 2016; Salvucci et 2016 Japan, Argentina ( Lindner 1887) Pediococcus lindneri silage, wheat and al. 2016) (Henneberg 1926) oat

*Lactobacillus kimchii is determined to be present in New Zealand under PNZ1000025 through the commercialisation of Kimchii (fermented vegetable) in New Zealand.

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Ubiquity of microorganisms

In 2014, EPA commissioned a report written by microbial ecologists from the University of Auckland to investigate the prospects around microbial ubiquity. The report discusses the theory and the extensive body of research supporting it that non-symbiotic or free-living prokaryotic microorganisms are ubiquitous in their distribution around the world and their growth is 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 by human vectors that have allowed non-symbiotic bacteria to disperse globally (Ehlers & Lear 2014).

This report has formed the basis of previous determinations on four nitrifying/denitrifying microbial species (APP201895), as well as for three different Lactobacillus species (APP203067). In both these determinations, the Decision-making Committee found that the species in question are geographically ubiquitous, based on their presence in a number of widely geographically separated regions. In addition, individual bacterial species are found in a variety of different environments. Therefore, the Committee considered that, on the balance of probability, these species did not meet the definition of a new organism as defined in the Act.

Like the bacteria in these decisions, the members of the order Lactobacillales under consideration in this determination have been isolated from various sources and used around the world to produce sourdough, alcohol, pickles, dairy products, and other foodstuffs. Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici are found in various fermentations (Yu et al. 2011; De Vuyst et al. 2014), contributing to the flavour and preservation of the food and beverages (Salvucci et al. 2016).

Products containing these bacteria, which include plants (Wratt & Smith 1983), cheese (Dacre 1958), yoghurt and meat, have been imported into or produced within New Zealand well before 29 July 1998. Moreover, the types of habitats where the ten organisms in this application are known to grow have been present in the New Zealand environment for a significant period of time, essentially coinciding with European colonisation of the country with the introduction of those fermented and dairy-based products by European settlers beginning in the late 18th century.

The ubiquity of Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici in geographically distinct environments internationally supports the conclusion that these species are likely to have been present in New Zealand prior to 29 July 1998, and to have an ongoing presence today.

EPA Staff Assessment Report: Application APP203045

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:

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 1.

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 Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici against this criterion.

Regarding other criteria listed in Section 2A of the Act, Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici:  have not been prescribed as risk species (section 2A(1)(b))  have not been approved to be held in containment or released with controls (sections 2A(1)(c), (ca) and (cb))  are not genetically modified organisms(section 2A(1)(d))  have 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 noted that they do not have in-house microorganism expertise and accordingly did not provide additional comment on this application. MPI did not provide any information or evidence that would

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help to determine whether Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici were present in New Zealand prior to 29 July 1998. 5. Overall findings and conclusion

Based on the information available, there is sufficient weight of evidence to conclude that Lactobacillus sanfranciscensis, L. rossiae, L. crustorum, L. curvatus, L. pontis, L. hilgardii, L. paralimentarius, Leuconostoc lactis, Pediococcus parvulus and P. acidilactici were likely to have been 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

Ahmadova A, Todorov SD, Hadji-Sfaxi I, Choiset Y, Rabesona H, Messaoudi S, Kuliyev A, de Melo Franco BDG, Chobert J-M, Haertlé T 2013. Antimicrobial and antifungal activities of Lactobacillus curvatus strain isolated from homemade Azerbaijani cheese. Anaerobe 20: 42-49. Arena ME, Lisi MS, Manca de Nadra MC, Alberto MR 2013. Wine composition plays an important role in the control of carcinogenic precursor formation by Lactobacillus hilgardii X(1)B. J Sci Food Agric 93(1): 142-8. Bull M, Plummer S, Marchesi J, Mahenthiralingam E 2013. The life history of Lactobacillus acidophilus as a probiotic: a tale of revisionary taxonomy, misidentification and commercial success. FEMS Microbiol Lett 349(2): 77-87. Cai Y, Okada H, Mori H, Benno Y, Nakase T 1999. Lactobacillus paralimentarius sp. nov., isolated from sourdough. International Journal of Systematic and Evolutionary Microbiology 49(4): 1451-1455. Cappelle S, Guylaine L, Gänzle M, Gobbetti M 2013. History and social aspects of sourdough. Handbook on Sourdough Biotechnology, Springer. Pp. 1-10. Corsetti A, Settanni L, van Sinderen D, Felis GE, Dellaglio F, Gobbetti M 2005. Lactobacillus rossii sp. nov., isolated from wheat sourdough. International Journal of Systematic and Evolutionary Microbiology 55(1): 35-40. Dacre JC 1958. 730. A note on the pediococci in New Zealand Cheddar cheese. Journal of Dairy Research 25(3): 414-417. Davis CR, Wibowo DJ, Lee TH, Fleet GH 1986. Growth and Metabolism of Lactic Acid Bacteria during and after Malolactic Fermentation of Wines at Different pH. Appl Environ Microbiol 51(3): 539-45. De Vuyst L, Van Kerrebroeck S, Harth H, Huys G, Daniel HM, Weckx S 2014. Microbial ecology of sourdough fermentations: diverse or uniform? Food Microbiol 37: 11-29. Douglas, H, Cruess, W 1936. A Lactobacillus from California wine: Lactobacillus hilgardii. Food Research 1: 113-119. Du Plessis H, Dicks L, Pretorius I, Lambrechts M, Du Toit M 2004. Identification of lactic acid bacteria isolated from South African brandy base wines. International journal of food microbiology 91(1): 19- 29. Ehlers C, Lear G 2014. The Biogeography of Environmental Microorganisms. Auckland Uniservices Limited. Elizaquível P, Pérez-Cataluña A, Yépez A, Aristimuño C, Jiménez E, Cocconcelli PS, Vignolo G, Aznar R 2015. Pyrosequencing vs. culture-dependent approaches to analyze lactic acid bacteria associated to chicha, a traditional maize-based fermented beverage from Northwestern Argentina. International Journal of Food Microbiology 198: 9-18. Garvie EI 1960. The genus Leuconostoc and its nomenclature. Journal of Dairy Research 27(2): 283-292. Giannino ML, Marzotto M, Dellaglio F, Feligini M 2009. Study of microbial diversity in raw milk and fresh curd used for Fontina cheese production by culture-independent methods. International Journal of Food Microbiology 130(3): 188-195. Giraffa G, Chanishvili N, Widyastuti Y 2010. Importance of lactobacilli in food and feed biotechnology. Research in Microbiology 161(6): 480-487. Gobbetti M, Minervini F, Pontonio E, Di Cagno R, De Angelis M 2016. Drivers for the establishment and composition of the sourdough lactic acid bacteria biota. International journal of food microbiology 239: 3-18. Gunther IL, White HR 1961. The Cultural and Physiological Characters of the Pediococci. Microbiology 26(2): 185-197. Heperkan D, Daskaya-Dikmen C, Bayram B 2014. Evaluation of lactic acid bacterial strains of boza for their exopolysaccharide and enzyme production as a potential adjunct culture. Process Biochemistry 49(10): 1587-1594. Kim HR, Lee AR, Kim JH, Ahn BH 2012. Microbial dynamics of commercial makgeolli depending on the storage temperature. J Microbiol Biotechnol 22(8): 1101-6. Kitahara M, Sakata S, Benno Y 2005. Biodiversity of Lactobacillus sanfranciscensis strains isolated from five sourdoughs. Letters in Applied Microbiology 40(5): 353-357. Lee J-S, Heo G-Y, Lee JW, Oh Y-J, Park JA, Park Y-H, Pyun Y-R, Ahn JS 2005. Analysis of kimchi microflora using denaturing gradient gel electrophoresis. International Journal of Food Microbiology 102(2): 143-150. Li Y, Wang F, Nishino N 2016. Lactic Acid Bacteria in Total Mixed Ration Silage Containing Soybean Curd Residue: Their Isolation, Identification and Ability to Inhibit Aerobic Deterioration. Asian-Australasian Journal of Animal Sciences 29(4): 516-522.

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Liu T, Li Y, Chen J, Sadiq FA, Zhang G, Li Y, He G 2016. Prevalence and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by culture dependent and pyrosequencing approaches. LWT - Food Science and Technology 68: 91-97. Lv X-C, Jia R-B, Li Y, Chen F, Chen Z-C, Liu B, Chen S-J, Rao P-F, Ni L 2016. Characterization of the dominant bacterial communities of traditional fermentation starters for Hong Qu glutinous rice wine by means of MALDI-TOF mass spectrometry fingerprinting, 16S rRNA gene sequencing and species-specific PCRs. Food Control 67: 292-302. Mamhoud A, Nionelli L, Bouzaine T, Hamdi M, Gobbetti M, Rizzello CG 2016. Selection of lactic acid bacteria isolated from Tunisian cereals and exploitation of the use as starters for sourdough fermentation. International Journal of Food Microbiology 225: 9-19. Muller MR, Ehrmann MA, Vogel RF 2000. Multiplex PCR for the detection of Lactobacillus pontis and two related species in a sourdough fermentation. Appl Environ Microbiol 66(5): 2113-6. Narváez-Zapata JA, Rojas-Herrera RA, Rodríguez-Luna IC, Larralde-Corona CP 2010. Culture-Independent Analysis of Lactic Acid Bacteria Diversity Associated with Mezcal Fermentation. Current Microbiology 61(5): 444-450. Osimani A, Garofalo C, Aquilanti L, Milanović V, Clementi F 2015. Unpasteurised commercial boza as a source of microbial diversity. International Journal of Food Microbiology 194: 62-70. Pepe O, Ventorino V, Cavella S, Fagnano M, Brugno R 2013. Prebiotic content of bread prepared with flour from immature wheat grain and selected dextran-producing lactic acid bacteria. Applied and environmental microbiology 79(12): 3779-3785. Pérez-Martín F, Seseña S, Palop ML 2015. Inventory of lactic acid bacteria populations in red wine varieties from Appellation of Origin Méntrida. European Food Research and Technology 240(4): 725-733. Sakai M, Nagano M, Ohta H, Kida K, Morimura S 2013. Isolation of Lactic Acid Bacteria from Takanazuke as a Starter Strain to Reduce Added Salt and Stabilize Fermentation. Food Science and Technology Research 19(4): 577-582. Salvucci E, LeBlanc JG, Pérez G 2016. Technological properties of Lactic acid bacteria isolated from raw cereal material. LWT - Food Science and Technology 70: 185-191. Scheirlinck I, Van der Meulen R, Van Schoor A, Huys G, Vandamme P, De Vuyst L, Vancanneyt M 2007. Lactobacillus crustorum sp. nov., isolated from two traditional Belgian wheat sourdoughs. Int J Syst Evol Microbiol 57(Pt 7): 1461-7. Sharafi H, Derakhshan V, Paknejad M, Alidoust L, Tohidi A, Pornour M, Hajfarajollah H, Zahiri HS, Noghabi KA 2015. Lactobacillus crustorum KH: Novel Prospective Probiotic Strain Isolated from Iranian Traditional Dairy Products. Applied Biochemistry and Biotechnology 175(4): 2178-2194. Sugihara TF, Kline L, Miller MW 1971. Microorganisms of the San Francisco sour dough bread process. I. Yeasts responsible for the leavening action. Appl Microbiol 21(3): 456-8. Vedamuthu E 1994. The dairy Leuconostoc: use in dairy products. Journal of Dairy Science 77(9): 2725- 2737. Viiard E, Mihhalevski A, Rühka T, Paalme T, Sarand I 2013. Evaluation of the microbial community in industrial rye sourdough upon continuous back–slopping propagation revealed Lactobacillus helveticus as the dominant species. Journal of Applied Microbiology 114(2): 404-412. Vogel RF, Böcker G, Stolz P, Ehrmann M, Fanta D, Ludwig W, Pot B, Kersters K, Schleifer KH, Hammes WP 1994. Identification of Lactobacilli from Sourdough and Description of Lactobacillus pontis sp. nov. International Journal of Systematic and Evolutionary Microbiology 44(2): 223-229. Weiss N, Schillinger U 1984. Lactobacillus sanfrancisco sp. nov., nom. rev. Systematic and Applied Microbiology 5(2): 230-232. Wirtz RL 2003. Grain, baking, and sourdough bread: A brief historical panorama. FOOD SCIENCE AND TECHNOLOGY-NEW YORK-MARCEL DEKKER-: 1-22. Wratt GS, Smith HC 1983. Plant Breeding in New Zealand. Elsevier Science. Yoon J-H, Kang S-S, Mheen T-I, Ahn J-S, Lee H-J, Kim T-K, Park C-S, Kho YH, Kang KH, Park Y-H 2000. Lactobacillus kimchii sp. nov., a new species from kimchi. International journal of systematic and evolutionary microbiology 50(5): 1789-1795. Yu J, Wang WH, Menghe BL, Jiri MT, Wang HM, Liu WJ, Bao QH, Lu Q, Zhang JC, Wang F and others 2011. Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia. J Dairy Sci 94(7): 3229-41. Zanirati DF, Abatemarco Jr M, Sandes SHdC, Nicoli JR, Nunes ÁC, Neumann E 2015. Selection of lactic acid bacteria from Brazilian kefir grains for potential use as starter or probiotic cultures. Anaerobe 32: 70-76.

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Appendix 1: 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 safekeeping facility’ and ‘immediately’ are defined in the Protocol Interpretations and Explanations of Key Concepts

3 Alternatively the application may lapse for want of information.

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EPA Staff Assessment Report: Application APP203045

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.

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