Electronic Appendix

Appendix 1. Invertebrate Surveillance System Components (SCC) and their definition.

SSC Definition of the SSC Pitfall traps are 43 mm diameter 100 ml vials containing 60 ml of ethylene glycol sunk to ground level. These traps rely on Pitfall traps (incl. targets falling in as they forage in the vicinity of the trap. A barrier trap is a pitfall trap set in the middle of a 2 m long fence of barrier pitfall traps) fly wire (10 cm high). The fence acts as a guide for walking insects to walk towards the trap. Barrier traps are run for 6 concurrent days, excluding the day of ‘rest’ following digging the trap in. 30 x 20 cm Perspex sheet is placed vertically over a plastic tray (35 x 25 cm) of soapy water. Flying insects fall into the water Window traps when they fly into the window. Window traps are left out for 48 hours. Water levels are checked after 24 hours and topped up if necessary. A dish with a small amount of each bait type (protein (e.g. tuna, egg) and carbohydrate (e.g. honey water, jam)) is left out for 2 Baited traps hours. Baits are checked every 20 minutes and ants in the vicinity of the baits are collected using either a wet paintbrush or soft forceps. Litter is scraped from a 1 x 1 m2 area, usually under trees and bushes. The litter is sieved to remove coarser material such as Litter extraction large twigs and rock then placed into a nylon sack. The litter is then transported back to the lab where it is placed into a Winkler sack. The Winkler sacks are run for four days with shaking of the litter occurring on the second day. Suction sampling 120 m2 area of shrubs are vacuumed with a motorised leaf vacuum/blower. 40 x 30 cm pine billets are placed on the ground weighted by a rock and labelled with a metal tag. The billet is left on the Wood baits ground for 6 months. Visual inspection of Inspections of wood structures, including natural wood, living or dead are carried out in natural vegetation or tenured areas. wood This SSC is usually done concurrently with Biologist structured surveys. Biologist structured A formal biological survey of a 10,000m2 area, focussing on areas in which exemplar species may be found. Specimens that survey cannot be easily recognized in the field as native are collected for identification by external subject matter experts.

Biologist The biologist undertaking surveillance on Barrow Island may detect a potential NIS in his/her personal space when travelling unstructured survey around the Island, but when not performing a Biologist structured survey.

Members of the island workforce who have not had any formal training to detect NIS (aside from the induction), but may Passive workers notice a potential NIS in their personal work or recreation area. Appendix 2. Vertebrate Surveillance System Components and their definition.

SSC Definition of the SSC Artificial habitat/refuge stations comprise horizontal stacks of alternating layers of treated timber, fibre cement sheeting (or similar) and local soil and plant material within a folded tarpaulin placed on Artificial habitat the ground, and held down by weights. Unbaited. Any indigenous vertebrate fauna utilising these stations is immediately removed and relocated at a sufficient distance to discourage return. One SSC is considered one Artificial habitat left in place for at least 4-6 weeks and then checked. These are 4-6 traps, spaced at approximately 10m intervals and placed in pairs on opposite sides of a single 60m length of 300mm tall flywire fence (remaining in situ for repeat sampling). Also deployed opportunistically along suitable artificial barriers such as fence lines and walls. If ambient Barrier funnel trap temperature, humidity and wind speed did not allow trapping during the heat of the day, traps were cleared of animals within three hours of sunrise, after which traps were closed and reopened in the afternoon within three hours of sunset. These traps are not baited. One SSC is considered one funnel trap left open for 24 hours and checked at least once. Pitfall traps were laid out in rows of 3-6 pits with up to 10m of fencing between traps. Traps left open for 7 to 14 days and checked daily. If ambient temperature, humidity and wind speed did not allow Barrier pitfall trap trapping during the heat of the day, traps were cleared of animals within three hours of sunrise, after which traps were closed and reopened in the afternoon within three hours of sunset. Traps not baited. One SSC is equivalent to 1 pit left out for one day of trapping.

A formal biological survey of an area looking for signs of non-indigenous vertebrate species. Signs may Biologist structured survey include tracks, scats, auditory calls, burrows, eye shine, eggs or individuals. One SSC is approximately 10,000m2 area.

An informal survey where animal signs are noted in the course of other activities (e.g. setting out Biologist unstructured survey traps). Where direct observation of a species is perhaps unlikely as the species is cryptic or nocturnal; scats, tracks, eggs, calls and burrows may be noticed by a biologist. A combination of small cages, rat tunnels and Tomahawk cages used. Traps baited using a combination of peanut butter and rolled oats. Traps were cleared three hours after sunrise, closed and then opened and rebaited (as required) after 1400 hrs daily. This avoided animals being re-trapped after release and Cage traps being confined during the heat of the day. A subset of cage traps were set on a raised table 122cm L X 60cm W x 73cm H with access to the table for target NIS (rodents) via two ramps, each 5cm wide and placed at an angle of 60 degrees (based on the methods of Morris 2002). One SSC unit equals 1 trap left out for one night of trapping. PCR Rodent WaxTag cards (Pest Control Research NZ). Traps baited using a combination of peanut butter and rolled oats. Placed and active for 7-14 day durations. Replaced as required. DNA from saliva deposited on the cards is sequenced to determine species identity (see methods). A subset of chew Chew cards cards were set on a raised table 122cm L X 60cm W x 73cm H with access to the table for target NIS (rodents) via two ramps, each 5cm wide and placed at an angle of 60 degrees (based on the methods of Morris 2002). One SSC equals 1 chew card left out for one night. Small Elliott type (Elliott Scientific Australia). Traps baited using a combination of peanut butter and rolled oats. Traps were cleared three hours after sunrise, closed and then opened and rebaited (as required) after 1400 hrs daily. This avoided animals being re-trapped after release and being confined Elliott traps during the heat of the day. A subset of Elliot traps were set on a raised table 122cm L X 60cm W x 73cm H with access to the table for target NIS (rodents) via two ramps, each 5cm wide and placed at an angle of 60 degrees (based on the methods of Morris 2002). One SSC unit equals 1 trap left out for one night of trapping. FADs consist of a wet tarpaulin or similar folded into layers and held down by bricks or rocks and Frog Attracting Device (FAD) placed in a shaded area. FADs are left out for a period of 2-3 nights, but checked every day for the presence of frogs. Not baited. One SSC is considered one FAD left out for one night.

Search of an area of likely habitat for the detection of invasive gecko species (vertical surfaces, near lights (attracted to invertebrate prey)). One SSC equals 1 light/wall surface searched for gecko scats. DNA from scat samples is sequenced to determine species identity (see methods). Sequences are Gecko scat analysis compared to taxa from the international sequence database (GenBank(ncbi.nlm.nih.gov/genbank/), the Helix vertebrate database and the Barrow Island specific database for Gekkonidae and Scincidae species. Hair Funnel (Faunatech Australia), baited with a combination of peanut butter and rolled oats. Placed and active for a 7-14 day duration; replaced as needed. Hairs captured in hair traps are analysed using genus-specific DNA probes developed for the identification of Rattus and Mus (see methods). A subset Hair traps of hair traps were set on a raised table 122cm L X 60cm W x 73cm H with access to the table for target NIS (rodents) via two ramps, each 5cm wide and placed at an angle of 60 degrees (based on the methods of Morris 2002). One SSC equals 1 hair trap left out for one night. Bushnell Trophy Cam Trail Cameras (Bushnell Australia), and ScoutGuard ZeroGlow 9M Cameras (ScoutGuard Australia). Active for 7-14 nights. Rebaited daily with a combination of peanut butter and rolled oats. Set to record in still (JPEG) or video (AVI) mode with a 30 second delay between successive Remote motion cameras recordings. A subset of remote cameras were set on a raised table 122cm L X 60cm W x 73cm H with access to the table for target NIS (rodents) via two ramps, each 5cm wide and placed at an angle of 60 degrees (based on the methods of Morris 2002). One SSC equals one nights recording on one camera. Sticky traps placed at likely habitats for the detection of gecko species. These include vertical surfaces near light sources. In order to reduce the risk of catching native species in this trap, the traps were only Sticky trap placed in environments that contain minimal numbers of native fauna (i.e. built environment). Traps were checked three hours after sunrise. This avoided animals being confined during the heat of the day. One SSC equals 1 sticky trap left out for one night and checked at least once.

Members of the island workforce who have not had any formal training to detect NIS (aside from the Passive workers induction), but may notice a potential NIS in their personal work or recreation area.