APPLICATION FORM Containment
To obtain approval for new organisms in containment
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
APP203908
Date
13 February 2019
www.epa.govt.nz 2
Application Form Approval for new organism in containment
Completing this application form
1. This form has been approved under section 40 of the Hazardous Substances and New Organisms (HSNO) Act 1996. It only covers importing, development (production, fermentation or regeneration) or field test of any new organism (including genetically modified organisms (GMOs)) in containment. If you wish to make an application for another type of approval or for another use (such as an emergency, special emergency or release), a different form will have to be used. All forms are available on our website. 2. If your application is for a project approval for low-risk GMOs, please use the Containment – GMO Project application form. Low risk genetic modification is defined in the HSNO (Low Risk Genetic Modification) Regulations: http://www.legislation.govt.nz/regulation/public/2003/0152/latest/DLM195215.html. 3. It is recommended that you contact an Advisor at the Environmental Protection Authority (EPA) as early in the application process as possible. An Advisor can assist you with any questions you have during the preparation of your application including providing advice on any consultation requirements. 4. Unless otherwise indicated, all sections of this form must be completed for the application to be formally received and assessed. If a section is not relevant to your application, please provide a comprehensive explanation why this does not apply. If you choose not to provide the specific information, you will need to apply for a waiver under section 59(3)(a)(ii) of the HSNO Act. This can be done by completing the section on the last page of this form. 5. Any extra material that does not fit in the application form must be clearly labelled, cross- referenced, and included with the application form when it is submitted. 6. Please add extra rows/tables where needed. 7. You must sign the final form (the EPA will accept electronically signed forms) and pay the application fee (including GST) unless you are already an approved EPA customer. To be recognised by the EPA as an “approved customer”, you must have submitted more than one application per month over the preceding six months, and have no history of delay in making payments, at the time of presenting an application. 8. Information about application fees is available on the EPA website. 9. All application communications from the EPA will be provided electronically, unless you specifically request otherwise.
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Commercially sensitive information
10. Commercially sensitive information must be included in an appendix to this form and be identified as confidential. If you consider any information to be commercially sensitive, please show this in the relevant section of this form and cross reference to where that information is located in the confidential appendix. 11. Any information you supply to the EPA prior to formal lodgement of your application will not be publicly released. Following formal lodgement of your application any information in the body of this application form and any non-confidential appendices will become publicly available. 12. Once you have formally lodged your application with the EPA, any information you have supplied to the EPA about your application is subject to the Official Information Act 1982 (OIA). If a request is made for the release of information that you consider to be confidential, your view will be considered in a manner consistent with the OIA and with section 57 of the HSNO Act. You may be required to provide further justification for your claim of confidentiality. Definitions
Restricting an organism or substance to a secure location or facility to prevent Containment escape. In respect to genetically modified organisms, this includes field testing and large scale fermentation
Any obligation or restrictions imposed on any new organism, or any person in relation to any new organism, by the HSNO Act or any other Act or any Controls regulations, rules, codes, or other documents made in accordance with the provisions of the HSNO Act or any other Act for the purposes of controlling the adverse effects of that organism on people or the environment
Any organism in which any of the genes or other genetic material: Have been modified by in vitro techniques, or Genetically Modified Are inherited or otherwise derived, through any number of replications, from Organism (GMO) any genes or other genetic material which has been modified by in vitro techniques
A new organism is an organism that is any of the following: An organism belonging to a species that was not present in New Zealand immediately before 29 July 1998; 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; An organism for which a containment approval has been given under the New Organism HSNO Act; An organism for which a conditional release approval has been given under the HSNO Act; A qualifying organism approved for release with controls under the HSNO Act; A genetically modified organism; An organism belonging to a species, subspecies, infrasubspecies, variety, strain, or cultivar that has been eradicated from New Zealand;
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An organism present in New Zealand before 29 July 1998 in contravention of the Animals Act 1967 or the Plants Act 1970. This does not apply to the organism known as rabbit haemorrhagic disease virus, or rabbit calicivirus A new organism does not cease to be a new organism because: It is subject to a conditional release approval; or It is a qualifying organism approved for release with controls; or It is an incidentally imported new organism
An individual or collaborative endeavour that is planned to achieve a particular Project aim or research goal
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Application Form Approval for new organism in containment
1. Applicant details
1.1. Applicant
Company Name: (if applicable) Manaaki Whenua - Landcare Research (MWLR)
Contact Name: Thomas Buckley
Job Title: Research Leader
Physical Address: 231 Morrin Road, St Johns, Auckland 1072
Postal Address (provide only if not the same as the physical): Private Bag 92170, Auckland Mail Centre, Auckland
Phone (office and/or mobile): + 64 9 574 4116
Fax:
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:
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2. Information about the application
2.1. Type of containment activity Tick the box(es) that best describe your application
Application type Type of new organism
☐ GMO Import into containment ☒ Non-GMO ☐ Develop in containment i.e. regeneration, fermentation GMO or genetic modification ☐ Non-GMO
☐ GMO Field test in containment ☐ Non-GMO
2.2. Brief application description Approximately 30 words about what you are applying to do
To import into containment, and hold in containment, New Caledonian stick insects of the order Phasmatodea to assess their thermal traits and forecast which species will survive climate change.
2.3. Summary of application Provide a plain English, non-technical description of what you are applying to do and why you want to do it
The purpose of this application is to import and hold foreign insects in the containment facilities of MWLR, to assess their thermal traits. In 2017, MWLR was awarded a Royal Society of New Zealand Marsden Fund grant to study how New Zealand stick insects have evolved from the warmer environments of their ancestral home in New Caledonia to withstand the temperate, variable environments of New Zealand to diversify into the 21 native New Zealand stick insect species present today. There are two possible mechanisms for a tropical insect to survive in a cooler, temperature environment - by tuning its biology to the environment or by broadening their baseline thermal preferences. Although studies of thermal preferences have been conducted in many insect species from a variety of environments, there are few studies that have examined these traits in an evolutionary context. We plan to do this by comparing thermal preferences between two well studied stick insect groups - the native New Zealand
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species to those of 4 related stick insect species from New Caledonia. This will help us understand whether tropical species that colonize temperate environments broaden their thermal preferences or rely on inherent ability to quickly regulate temperature relevant traits. Understanding these mechanisms and thermal preferences of New Zealand and New Caledonian stick insects in this way will also ultimately improve forecasts for which insect species here in New Zealand and in New Caledonia will be vulnerable to climate change. If these stick insect species are not able to quickly regulate their thermal preferences when put into a new thermal environment, this will greatly reduce survival of these species if the pace of climate change outpaces their ability to broaden their thermal preferences over time.
MWLR has two MPI accredited containment facilities that make it possible to investigate the activity of non-native invertebrates without risk to native New Zealand species and populations. Insects will only leave the containment facility after being killed via flash freezing in liquid Nitrogen and stored in an ultra-freezer. We are confident that flash freezing will kill these species as we know that the New Zealand species cannot tolerate temperatures below -10 °C (Dennis et al. 2015).
To assess the thermal preferences of New Caledonian stick insect species, these species need to be brought back to MWLR facilities and allowed to adjust to lab conditions for 7 days. After this adjustment period is over, insects will be divided into one of two temperature exposures – a temperate, 10C exposure and a warmer, 20C exposure using environmental chambers that can maintain these temperatures consistently for a set period. After a week has passed at these temperatures, we will then quantify how much each individual stick insect consumes and excretes during a 12 hour, overnight feeding trial at 8 different test temperatures ranging from 0 to 35C. The total trial length is approximately 4.5 weeks from arrival in the Containment Facility to completion of the experiments.
These trials will allow us to visualize a curve and determine at which temperatures the insects are most active. Experiments during 2018 and 2019 with New Zealand species found that the best way to make sure stick insects actually eat and excrete during these tests involves a 36 hour starvation period between each overnight trial, thus resulting in a 16 day experiment in addition to the two weeks of lab adjustment and temperature exposure for a total of 30 experiment days. Our experiments with the New Zealand species also indicate that females are best to quantify feeding rate as they are generally larger than the males, therefore eating more, and the need for nitrogen for egg production ensures that they are actively feeding while in captivity. The ability to import these New Caledonia species will improve the quality of our data set by allowing us to use proper lab equipment at MWLR when collecting our data as appropriate lab facilities in New Caledonia are lacking. It will also greatly reduce costs of conducting such a long experiment by decreasing the costs of lodging and shipment of necessary scientific
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equipment and other supplies while working abroad thus allowing for a better use of Marsden Funds.
We propose to collect nymphs and adult females from described and undescribed species of New Caledonian stick insect. These are all new species to New Zealand and require EPA approval. We will not be importing these live organisms on any plants as the New Caledonia stick insects will be able to survive during transportation for up to 24 hours without food and once brought into New Zealand can survive on locally grown and collected Mestrosideros excelsa and Rubus sp. brambles as they feed on close relatives of these plant species in New Caledonia. We plan to import these species in 3-layers of protective packaging consisting of individually packing each stick insect collected in New Caledonia into a 50ml Falcon tube with screw top lid, placing these tubes inside larger plastic, containers with snap lids, and then storing these larger, snap lid containers in a chilly bin sealed with duct tape. Female insects may lay eggs during transport, so we may also import eggs but we do not intend to rear these species in containment and therefore any eggs that are laid will be flash frozen once they have arrived into containment or laid during the experiment period.
2.4. Background and aims of application
This section is intended to put the new organism(s) in perspective of the wider activities that they will be used in. You may use more technical language but all technical words must be included in a glossary
The purpose of this application is to import and hold in MWLR’s Beever Plant Pathogen Containment facility (Facility) foreign insects in order to understand their thermal preferences and forecast which species will be able to survive climate change.
Temperature, more than any other factor, governs the distribution of plants and animals. Species’ traits are fine-tuned to a given temperature regime – their thermal performance. Revealing the processes that determine this thermal performance is therefore the key to understanding how organisms have evolved to occupy new thermal environments. Many insect lineages evolved in the tropics where temperatures are warm and stable, and a narrow thermal performance is presumed advantageous for these species. However, temperate environments tend to be more thermally variable as well as cooler. Therefore, if a tropical lineage is to survive and reproduce when introduced into a temperate environment, it must either broaden its baseline thermal performance or rely on plasticity to tune its thermal performance to the environment. Surprisingly, although there are many single-species studies of thermal performance in different environments, this assumption has not been thoroughly tested in an evolutionary context. We therefore cannot determine whether tropical species that colonize temperate environments broaden their thermal performance or rely on their inherent plasticity. This fundamental
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question must be resolved to advance our understanding of evolutionary diversification and biogeographic patterns. The integration of genomics, physiology, and phylogeny now allows us to address this question using the well-studied clade of New Zealand and New Caledonian stick insects.
A thermal performance curve (TPC) describes how the performance of an organism varies over a range of temperatures in a quantitative manner by measuring how much an individual “performs” at a range of different temperatures. Performance can be quantified by measuring any biological process that is temperature dependent which includes consumption, excretion, mating, egg production, walking etc. in insects because they are ectothermic and therefore their body temperatures reflect the temperature of their environment. Over evolutionary time, a species can broaden or shift its TPC. For example, in order for the first stick insects that arrived in New Zealand from New Caledonia to have survived, the stick insects could have broadened their TPCs from a narrow curve that included only warm, hot temperatures that they were exposed to in New Caledonia to include cold temperatures experienced in New Zealand OR they could have shifted their TPCs to no longer include hot temperatures in favour of colder temperatures. For the last year, we have been conducting experiments with New Zealand stick insects species to understand which species have broad or narrow TPCs and how they shift when exposed to a cold or warm temperature for an extended period (ie. Plasticity). Preliminary findings suggest that northern species have narrower TPCs, some mid-latitude species are plastic and are able to shift their performance after a cold or warm exposure, and southern species have broader TPCs allowing them to function at both cold and hot temperatures to survive in the variable climates of the South Island. By conducting the same experiments in ancestral New Caledonian stick insects species, we will be able to understand how the thermal traits of these New Zealand species have evolved after introduction to colder, more variable New Zealand environments and better understand how the environment has diversified New Zealand stick insects into the 21 species recognized today. Understanding how these thermal traits have evolved in these species will allow us to better forecast which of these species as well as other insect groups in New Zealand will be able to survive climate change.
Lab facilities in New Caledonia do not have the proper number of environmental chambers necessary in order to answer these questions about thermal preferences in New Caledonia stick insect species. Conducting our experiment there will limit the number of stick insect species and the number of individuals per species that we would be able to study, greatly decreasing the statistical power and scope of our research project. Assessment of these traits will be greatly improved by conducting experiments in the MWLR containment facility in Auckland. MWLR’s MPI accredited Transitional and Containment facility for the containment of plants, invertebrates and microbes based in Auckland at the MWLR Tamaki campus makes it possible to investigate the thermal traits
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of New Caledonian stick insect species without the risk of escape into the environment. The facility is certified for the testing of rust fungi and exotic invertebrates, and to house exotic plants.
3. Information about the new organism(s)
3.1. Name of organism Identify the organism as fully as possible
Non-GMOs - Provide a taxonomic description of the new organism(s).
GMOs – Provide a taxonomic description of the host organism(s) and describe the genetic modification.
Both - Describe the biology and main features of the organism including if it has inseparable organisms. Describe if the organism has affinities (e.g. close taxonomic relationships) with other organisms in New Zealand. Could the organism form an undesirable self-sustaining population? If not, why not? How easily could the new organism be recovered or eradicated if it established an undesirable self- sustaining population?
Identification of the Organisms: Endemic species of Phasmatodea from New Caledonia including Canachus alligator, Carlius fecundus, Cnipsus rachis, Leosthenes aquatilis, Aprenas effeminatus, Asprenas impennis, Microcanachus matileorum, Labiodiophasma rouxi, Caledoniophasma marshallae, Trapezaspis pinoruminsulae, undescribed species of the above genera (see Buckley et al. 2010) and undescribed genera (see Buckley et al. 2010) Type of organism (eg bacterium, virus, Animal fungus, plant, animal, animal cell): Taxonomic class, order and family: Asprenas effeminatus (Carl, 1913) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Asprenas impennis (Carl 1915) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
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Canachus alligator (Redtenbacher, 1908) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Carlius fecundus (Carl, 1915) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Cnipsus rachis (Saussure, 1868) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Leosthenes aquatilis (Stal, 1875) – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Microcanachus matileorum Donskoff, 1988 – Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Labiodiophasma rouxi (Carl 1915) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Caledoniophasma marshallae (Zompro, 2001), Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Trapezaspis pinoruminsulae (Delfosse, 2013), Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Nov. Gen. 1, nov. sp. 1 (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Nov. Gen. 2, nov. sp. 1 (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Nov. Gen. 2, nov. sp. 2 (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Asprenas sp. (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
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Canachus sp. (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Carlius sp. (see Buckley et al. 2010) - Insecta (Class), Phasmatodea (Order), Phasmatidae (Family)
Describe the biology and main features of The New Caledonian stick insects are all the organism including if it has inseparable plant feeding species that inhabit forest organisms. and scrub. They have been collected from a wide range of New Caledonian plants including from the familied Myrtaceae, Cunoniaceae and various ferns. They are usually nocturnal feeders. Males and females are known from all the described New Caledonian species suggesting they reproduce sexually unlike some other stick insect species. The females lay eggs which are randomly dropped. It is unknown how long the egg develpment time is but in most other stick insects species it is in the order of months. Most species are flightless where the wings have been reduced to small, vestigial flaps involved in either predator defense or mate signalling. Two genera do include fully flighted species; Leosthenes and “Nov. Gen. 2” from Buckley et al. (2010). The smallest species, Microcanachus matileorum, is approximately 2.5 cm long when fully grown. The largest species are from the genus Asprenas and the females are approximately 10 cm long. Stick insects fom other countries can be infected with Mermithidae nematodes (Yeates and Buckley 2006). Occasionally mites can be seen on the stick insect exoskeleton. It is unknown if this is the case for New Caledonian species.
Morphological information on the New Caledonian stick insects is relatively scarce but can be summarized below for the genera:
Asprenas: These are relatively large insects with the females over 10 cm for some species
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and the males a bit shorter than this. They tend to be mottled brown, grey or light green. They usually have spines on the dorsal surface. They lack functional wings. Canachus: These are moderate to large, robust insects and the females are between 5 and 10 cm. They can have spines on the dorsal surface and lack functional wings. They are usually brown in colour. Cnipsus: These are small insects, less than 5 cm and have relatively long legs. They often are found on ferns. They are usually brown or green and have slender spines on the dorsal surface. Carlius: These are long, gracile insects and lack wings or spines. They can be brown or green. Leosthenes: The males and females are flighted and are slightly less than 5 cm to approximately 7 cm in length. They are mottled and usually grey or brown in colour. Caledoniophasma: These are robust, broad insects and are brown in colour. Females are a bit over 5 cm in length and males somewhat shorter than this. Very few specimens are known. They are brown in colour. Microcanachus: These are very similar to Canachus except they are short in size, being 2.5 cm (females) or less (males). They seem to inhabit leaf litter. Trapezaspis: These are less then 4 cm long and are robust, broad insects. They have no functional wings and are brown or grey in colour. They often have spines on the thorax. Labiodiophasma: Similar to Trapezaspis, these insects are approximately 5 cm long or shorter in the case of males. They have prominent spines on the thorax and are typically brown. Nov. Gen. 1: These are the most distinctive species. They are about 5 cm long, and the males are a bit shorter. They are brightly coloured with various colour morphs observed. These are usually combinations or pink and black, yellow and black or red/orange and black. They are not flighted. Nov. Gen. 2: These are very gracile insects, more so than any other New Caledonian species. They are flighted and
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are usually brown, grey or green in colour. The females are approximately 7 cm in length and the males shorter.
Describe if the organism has affinities (e.g. All of the New Caledonian stick insects close taxonomic relationships) with other form a monophyletic group that is closely organisms in New Zealand. related to the New Zealand stick insects (see Buckley et al. 2010).
Could the organism form an undesirable Climate matching can be used to identify self-sustaining population? If not, why regions that could potentially be colonized not? by an introduced species on the basis of similarity to climates found in the species’ native range. However, we cannot produce detailed climate match models due to a lack of stick insect physiological data to create such models for the species we intend to import. Auckland is a sub- tropical environment so it is likely that New Caledonia species could establish in the unlikely event they escaped containment because Auckland does not experience hard freezes in the winter months. However, if potentially introduced species’ moved south – it would likely become impossible for these populations to establish in colder, more variable environments as these cold temperatures would not be common within New Caledonia. It is also likely that the New Caledonian species would be able to feed on New Zealand species of Myrtaceae plants as they feed on this family on New Caledonia. How easily could the new organism be It should be possible to locate and eradicate recovered or eradicated if it established an a self-sustaining population outside the undesirable self-sustaining population? containment facility if detected early. Because our experimental protocol demands we track each individual on a daily basis, we will quickly be able to notice if one of our specimens goes missing. In the very unlikely event that a misplaced specimen escapes from the environmental chamber, we are confident we would be able to find the missing organism before it was able to establish a self-sustaining population. The stick insects species we propose to import range in size from 2.5 to 10 cm and are therefore easily visible within the laboratory environment and for trained personel outside of the lab. Their size though greatly
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increases the chance that a stick insect would be found before it could escape from containment. Most of the species we will import are flightless. Importing only females and nymphs greatly reduces the risk of a self-sustaining population establishing.
3.2. Regulatory status of the organism
Is the organism 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
4. Information about the containment
4.1. For field tests: The nature and method of the field test Describe the nature and method of the field test and the experimental procedures to be used
N/A
4.2. Proposed containment of the new organism(s) (physical and operational)
Describe how you propose to contain the new organism(s) after taking into account its ability to escape from containment (i.e. the possible pathways for escape)
The insects will be contained in MPI accredited PC2 facilities designed, built and actively managed to contain plants, invertebrates and micro-organisms (5713 / ATP 26909). Containment in these facilities is achieved by a combination of the strict laboratory operating procedures and the physical specifications of the building.
Theoretically, escape of the insects from containment could occur:
1. By plant or insect material being inadvertently carried out of the facility by personnel on contaminated clothing or equipment.
2. In association with water movement of eggs that were laid during containment.
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3. During waste disposal of insect or plant material.
4. In transit during transfer to, or from, the facility.
5. By being deliberately removed from the facility by unauthorised persons.
To address these risks:
• Access to the Facility is restricted by swipe card access to trained, authorised personnel only. This includes maintenance contractors who must also be trained in containment procedures prior to being accompanied in- and out of the building.
• The Facility is designed and built to comply with Physical Containment level 2 (PC2) AS/NZS 2243.3.2002: Safety in Laboratories part 3, and to the MAF standards 154.03.02 Microorganisms and Cell Cultures: 2007a, 154.02.08, “Transitional and Containment Facilities for Invertebrates, and Transitional and Containment facilities for Plants 2007. In addition, there are quality assurance systems in place for the Facility to achieve a level of physical containment that exceeds level PC2 and the MAF Standards. Additional containment features include; 1) directional negative air flow within the building leading to the corridor and wet lab areas, 2) HEPA filtration of all air exiting the building, 3) closed air recirculation systems for each of the high level containment growth rooms, 4) heat sterilisation treatment of all waste water prior to it leaving the building, 5) decontamination of all equipment and clothing leaving the building and 6) the showering of personnel who have entered the containment rooms prior to leaving the building. These will be explained in more detail below.
• Containment within the Facility is based on a four level of sealed compartments system which has essentially four levels of sealed compartments between the highest containment area (i.e. growth rooms and laboratory) and the outside. There are four doors separating each growth room and the laboratory from the outside representing four levels of containment. Each level is monitored for the possibility of escape from the previous level with the use of sticky traps designed to trap insects, pollen, air borne seeds and spores, which are checked regularly. There are procedures outlined in the facility manual (Appendix 10) should an escape be detected at any level, to prevent further escapes and to reduce the chances of the escapee from making it outside.
• The Facility is mechanically ventilated to ensure a directional air-flow is maintained. Growth-room air is extracted initially through a wire mesh designed to capture all potential invertebrate vectors. Each growth room has a closed air recirculation system to ensure there is no cross contamination between rooms and
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re-circulated air is not released outside unless vented through a HEPA filter which filters the air at the micron level capturing all spores, pollen dust particles, etc. There are no opening external windows in the facility.
• The Facility has a negative air pressure gradient that ensures all air flows towards the inner corridor and wet laboratory areas ensuring no pollen, air borne seed, dust or potential invertebrate vectors can escape the facility when doors are opened.
• To ensure invertebrates and eggs are not inadvertently carried out of the Facility by personnel, staff are required to wear coveralls, hair covers, shoe covers, and gloves. When exiting the facility, staff must remove any coveralls, hair covers, shoe covers, and gloves, and wash their hands and fingernails after working with these organisms before they leave the Facility. Those staff that have entered Growth Rooms are required to decontaminate via showering.
• Entrance into the Facility is via magnetic card swipes, that are only issued to certified facility users, and an airlock to ensure any airborne pollen, seeds and invertebrates are contained within the facility. There is also a light trap inside the air lock to trap any insect that may make it as far as the air lock (or that enter from the outside).
• Plants will be grown and maintained in the dry lab of Facility and at the end of an experiment the room will be cleared of all materials, cleaned and decontaminated according to the procedure outlined in the Facility manual.
• All excess plant material, biological and organic waste, coveralls, hair covers, shoe covers, and gloves are autoclaved at 121 °C and 15 psi for 90-minutes and then disposed of via Interwaste Secure Disposal, as described within the Facility operation manual.
• No viable propagules or biological material will be removed from the laboratory unless via an approved Movement Request (MPI Containment Verifier) to be transported to an appropriate containment facility or shipped overseas to a suitable facility or for release. All organisms moved into/out of the Facility to a remotely located facility will be double packaged and transported from the laboratory in accordance with Packaging Instruction No. 650 of the IATA) Dangerous Goods Regulations. This involves shipping plants and biocontrol agents in plastic containers placed inside zip-lock bags which are then put in an ice box which is sealed shut using packaging tape.
• All liquid waste, including cleaning waste, flows into a 1000 L holding tank. The tank is secondarily contained in a 1000 L, bitumen-lined, concrete water tank. The tank contents are then heat treated to 125°C for 30-minutes, 20 L at a time. The
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tank contents are then heat treated to 125°C kill any viable propagules on a weekly basis or more regularly if required. Following treatment the waste is cooled to 40ºC, before being discharged to the sanitary sewer (Appendix 9c, Facility Containment Manual, Sterilised liquid waste disposal).
• All solid waste including coveralls, soil, plant and microbial materials are autoclaved to 121 °C and 15 psi for 90-minutes within the wet laboratory before exiting the Facility for disposal via Interwaste certified disposal.
• Any equipment leaving the Facility is decontaminated with either 70% ethanol spray and/or 2% Sterigene solution following the appropriate procedures outlined in the facility manual to ensure no viable plant material leaves the Facility.
• Following the experiments the insects are killed by immersion in liquid nitrogen and then transferred from the Facility and into an ultrafreezer at Manaaki Whenua.
• We will freeze frass samples collected during the experiments at -20 °C. The frass will then be transported out of the containment facility to enable biochemical analysis of proteins and other macromolecules. It is straightforward to identify eggs amongst frass and remove them for descruction. On the off chance any eggs are not removed from the frass, the treatment at -20 °C will render them non- viable.
In the unlikely event of an incident or accident that may lead to an escape, the PC2 transitional and containment Facility has an organism escape contingency plan (refer to the Facility Manuals). This involves notifying the MPI Containment Verifier as soon as possible and within 24 hours of noticing breach of containment. The MPI Containment Verifier will then decide on a course of action depending on their assessment of the risk.
5. Māori engagement
Discuss any engagement or consultation with Māori undertaken and summarise the outcomes. Please refer to the EPA policy ‘Engaging with Māori for applications to the EPA’ on our website (www.epa.govt.nz) or contact the EPA for advice.
Consultation with local Māori groups (namely Ngati Paoa, Ngai Tai Ki Tamaki, Ngāti Whātua Ōrākei and Waikato Tainui) took place on the 9th of October via e-mail. One reply was received from Ngati Paoa who expressed no problem with importing into containment only (see Appendix 3).
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6. Risks, costs and benefits
Provide information of the risks, costs and benefits of the new organism(s).
These are the positive and adverse effects referred to in the HSNO Act. It is easier to regard risks and costs as being adverse (or negative) and benefits as being positive. In considering risks, cost and benefits, it is important to look at both the likelihood of occurrence (probability) and the potential magnitude of the consequences, and to look at distribution effects (who bears the costs, benefits and risks).
Consider the adverse or positive effects in the context of this application on the environment (e.g. could the organism cause any significant displacement of any native species within its natural habitat, cause any significant deterioration of natural habitats or cause significant adverse effect to New Zealand’s inherent genetic diversity, or is the organism likely to cause disease, be parasitic, or become a vector for animal or plant disease?), human health and safety, the relationship of Māori to the environment, the principles of the Treaty of Waitangi, society and the community, the market economy and New Zealand’s international obligations.
You must fully complete this section referencing supporting material. You will need to provide a description of where the information in the application has been sourced from, e.g. from in-house research, independent research, technical literature, community or other consultation, and provide that information with this application.
The potential risks, costs and benefits of the proposed introduction to NZ containment of the New Caledonian stick have been identified by literature review and our own knowledge of the biology and ecology of New Zealand and New Caledonian stick insects. All effects considered to be potentially significant are addressed in detail.
The main risks are associated with the potential for imported stick insects to ‘escape’ containment and established feral populations. Potential costs are associated with eradicating insects if a breach from containment occurs. Benefits of importing plants are scientific.
Risks
Potential adverse effects on the environment, in particular on ecosystems and their constituent parts could only occur if imported insects ‘escaped’ containment and establish feral populations. As noted in Section 4.2. there are many measures in place to prevent this. In the unlikely event that an imported stick insect escapes and establishes in NZ possible adverse effects could be:
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Deterioration of ecosystems and loss of biodiversity, (e.g. by all listed insects indiscriminately outcompeting/smothering native species) if the insects cannot be eradicated from the environment.
Potential impacts on the health of our native and valued fauna. Although, to our knowledge, none of the listed species are known to be toxic or allergenic to animals or humans.
Likely flow-on effects to the mauri (life essence) of those species and the role of Māori as kaitiaki (guardians/stewards) in the maintenance and management of mauri.
A breach of containment enabling an insect to ‘escape’
All MWLR’s Beever Plant Pathogen Containment facility (Facility) users must undertake training before being allowed to work in the facility and will be aware that no organism can be deliberately taken out of the facility without the appropriate approvals.
The risk that insects will be accidentally taken out of containment is extremely low:
It is possible that insects could adhere to clothing, but all staff in the facility wear disposable coveralls, gloves, hair nets and disposal boot-covers, that must be removed and disposed of, prior to leaving the building. In addition, because our experiments require the tracking of individuals over time, each individual insect imported into NZ and housed in containment will be housed in its own, 1L container and checked daily so it will be easy to keep track of each individual and constantly be vigilant about potential escapees. If a stick insect becomes misplaced in the containment facility, it will be noticed quickly and found easily as the size of all the stick insects we will be importing range in length from about 2.5 cm to over 10 cm in length and therefore easily identifiable by eye alone.
Two genera contain flight capable species. Flying insects tend to fly towards bright light, so if they escape from the rearing container/temperature chamber then they are likely to fly to the window in the dry laboratory (where they can easily be detected/trapped), rather than towards the corridor. If they did somehow escape into the corridor/wet lab area, again they are most likely to orient towards the window. It is highly unlikely that they would find their way out of the containment building as this would require passing through a minimum of 3 more doors: one into the change room and then two more (the double door entry – which is an airlock) to reach the outside. In addition, a light trap operates continually between the doors of the double-door ‘airlock’ system which should kill any flying insects that make it into the airlock (the only entry/exit from the containment facility) and sticky traps are suspended throughout the facility to trap flying insects.
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Insect establishment in NZ
The stick insects are capable of feeding on plant species that are present in New Zealand. Establishment of stick insects locally is therefore a risk as there are local food sources available, but we will minimize this risk by importing only females of these species. If females do not have access to males, this will greatly decrease the risk of establishment as they won’t be able to successfully mate and reproduce in the wild in the highly unlikely event of insects escaping containment. However it is possible that the adult females we collect will already have mated in New Caledonia and so may contain fertilized eggs. To become established would require insects to grow and reproduce outside containment without being detected until eradication is no longer feasible. Detection of insect populations near the facility is highly likely (the surrounding areas are used by MWLR and MPI staff to propagate plants).
The New Caledonian and New Zealand stick insects diverged approximately 15 million years ago (Simon et al. 2019). Therefore their genomes are likely to have diverged sufficiently to make hybridisation impossible.
Cost
This is hard to estimate but if we assume an affected area of one hectare then the total cost of eradication may be less then $100,000 following Brockerhoff et al. (2010).
Benefits
New Zealand is unique in being one of the most isolated, ancient, and topographically diverse continental islands resulting in a high richness of endemic species, the great majority of which are invertebrates. However, the effects of climate change on native New Zealand invertebrates have not been extensively studied. This research into understanding how tropical lineages are able to maintain fitness in a new environment will be beneficial in predicting which NZ species will survive climate change by furthering our understanding of the biological strategies these insects employ to withstand environmental change. Although climate change effects on New Zealand insects are not well studied, a study looking at climate change effecs on alpine plant biodiversity suggests that if present mean temperatures of ~0.6C higher than in 1900 are maintained in combination the threat of exotic species introduction, 40-70 native plants will become at risk of extinction. Many New Zealand stick insects are cold adapted and therefore also at risk.
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Being able to control the quality of the research undertaken to assess risk vs benefit leading to even safer releases of biological agents to control weeds in countries that are unable to conduct such assessments as they do not have expensive containment facilities.
The opportunity to further increase the skills and knowledge of NZ researchers through the opportunity to work on a wider range of targets, and interact with international researchers, so that we are better prepared to defend the NZ environment from future insect invasions.
Undertaking this work for New Caledonia will also facilitate future opportunities for NZ scientists to participate in other projects in the future, designed to conserve the natural environment and improve productivity of poorer countries with limited resources to do so.
7. Alternative methods and potential effects from the transfer of genetic elements This section is for developments of GMOs that take place outdoors and field tests of GMOs only
Discuss if there are alternative methods of achieving the research objective. Discuss whether there could be effects resulting from the transfer of genetic elements to other organisms in or around the site of the development or field test.
N/A
8. Pathway determination and rapid assessment This section is for the imports of GMOs only
Under section 42B of the HSNO Act your application may be eligible for a rapid assessment. The pathway for your application will be determined after its formal receipt, based on the data provided in this application form. If you would like your application to be considered for rapid assessment (as per the criteria below), we require you to complete this section.
8.1. Discuss whether the GMO(s) to be imported fulfil the criteria
The criteria are: The host organism(s) are Category 1 or 2 host organisms as per the HSNO (Low Risk Genetic Modification) Regulations The genetic modifications are Category A or B modifications as per the HSNO (Low Risk Genetic Modification) Regulations and the modifications are not listed in the Schedule of these Regulations The minimum containment of the GMO(s) will be as per the HSNO (Low Risk Genetic Modification) Regulations (PC1 or PC2 as per AS/NZS2243.3:2002)
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N/A
9. Other information
Add here any further information you wish to include in this application including if there are any ethical considerations that you are aware of in relation to your application.
In addition to this EPA approval, import approvals from MPI will be required for each individual insect species to be imported. Collection permits for use in New Caledonia will also be obtained.
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10. 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 n/a identified appendix
Supplementary optional information attached:
Copies of additional references ☒ Yes ☐ No Relevant appendices attached
Relevant correspondence ☒ Yes ☐ No See appendix
Administration Are you an approved EPA customer? ☒ Yes ☐ No I assume MWLR is approved 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
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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.
Thomas Buckley
30/10/2019
Signature Date
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:
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Appendices and referenced material (if any) and glossary (if required)
APPENDIx 1 1: Manaaki Whenua Landcare Research Beever Plant Pathogen Containment Facility Manual See attached file. We request that the contents of this manual remain confidential.
APPENDIX 2: References
Brockerhoff, E.G., A.M. Liebhold, B. Richardson, and D.m. Suckling. 2010. Eradication of invasive forest insects: concepts, methods, costs and benefits. New Zealand Journal of Forestry Science, 40 Supp., S117-S135.
Buckley, T.R., D. Attanayake, J.A.A. Nylander, and S. Bradler. 2010. The phylogenetic placement and biogeographical origins of the New Zealand stick insects (Phasmatodea). Systematic Entomology, 35: 207–225.
Dennis, A.B., L.T. Dunning, B.J. Sinclair, and T.R. Buckley. 2015. Parallel molecular routes to cold adaptation in eight genera of New Zealand stick insects. Scientific Reports, 5: 13965.
Simon, S., H. Letsch, S. Bank, T.R. Buckley, S. Liu, R. Machida, B. Wipfler, X. Zhou, and S. Bradler. 2019. Old World and New World Phasmatodea: Phylogenomics resolve the evolutionary history of stick and leaf insects. Frontiers in Ecology and Evolution, in press.
Yeates, G.W., and T.R. Buckley. 2009. First records of mermithid nematodes (Nematoda: Mermithidae) parasitising stick insects (Insecta: Phasmatodea). New Zealand Journal of Zoology, 36: 35-39.
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APPENDIX 3: E-mail sent to Ngati Paoa, Ngai Tai Ki Tamaki, Ngāti Whātua Ōrākei and Waikato Tainui
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Replies from mana whenua
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