Mesa a Hub: Warramboo Troglobitic Fauna Assessment

Home , Draculoides, Draculoides bramstokeri, Schizomida, Stricticollis, Stygiocaris, Zygentoma

Mesa A Hub: Warramboo Troglobitic Fauna Assessment

Prepared for Rio Tinto

July 2017 Warramboo Troglofauna Assessment

This page intentionally blank

2 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Job No.: 1080

Prepared by: Jason Alexander

Document Quality Checking History Version: Rev A Peer review: Sylvie Schmidt Version: Rev 0 Director review: Garth Humphreys Version: Rev 0 Format review: Fiona Hedley

Approved for issue: Garth Humphreys

This document has been prepared to the requirements of the client identified on the cover page and no representation is made to any third party. It may be cited for the purposes of scientific research or other fair use, but it may not be reproduced or distributed to any third party by any physical or electronic means without the express permission of the client for whom it was prepared

or Biota Environmental Sciences Pty Ltd.

This report has been designed for double-sided printing. Hard copies supplied by Biota are printed on recycled paper.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 3 Warramboo Troglofauna Assessment

This page intentionally blank

4 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Warramboo Troglofauna Assessment

Contents

1.0 Project Summary 9 2.0 Executive Summary 11 3.0 Introduction 13 3.1 Study Objectives and Scope 13 3.2 Purpose of this Report 13 3.3 Background on Subterranean Fauna 15 3.4 Terminology 16 4.0 Methodology 17 4.1 Desktop Review and Database Searches 17 4.2 Habitat Characterisation 17 4.3 Sampling Methodology 18 4.4 Molecular Analysis 20 4.5 Categories of Conservation Significance 20 4.6 Survey Design 21 4.7 Limitations 25 5.0 Results 27 5.1 Study Area Habitat Characterisation 27 5.2 Desktop Review Results 35 5.3 Survey Results 43 6.0 Discussion 55 6.1 Recorded Troglobites Excluded from Assessment 55 6.2 Conservation Significance 55 6.3 Habitat Suitability 59 6.4 Conclusions 62 7.0 Glossary 63 8.0 References 65

Appendix 1 Licence to take Fauna for Scientific Purposes (licence: SF010296) Appendix 2 Raw Sampling Data Appendix 3 Desktop Review: Troglobitic Fauna Species Lists and Conservation Significance Appendix 4 Helix Molecular Solutions:

Genetic Analysis Reports Appendix 5 WAM Short-Range Endemic Categories Summary

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 5 Warramboo Troglofauna Assessment

Appendix 6 Desktop Review: Previous Studies and Comparison

Tables Table 1.1: Summary of project specifications and limitations. 9 Table 4.1: Summary and location of database searches completed for the current scope 17 Table 4.2: Conservation classification used within this report. 20 Table 4.3: Summary of field mobilisations completed for the Warramboo Troglofauna survey area. 21 Table 4.4: Name, location and sampling information of drill holes sampled for troglofauna within the Warramboo survey area. 22 Table 5.1: Surface geology of the Warramboo troglofauna study area and potential for occurrence of subterranean fauna. 27 Table 5.2: Prospectivity of the geological units within the study area to provide troglofauna habitat. 33 Table 5.3: Description of PECs overlapping the study area. 35 Table 5.4: Summary of previous surveys completed within the survey area and surrounds. 38 Table 5.5: Taxa and abundance (n) of troglofauna recorded from the desktop review. 40 Table 5.6: Order-level identifications and sequencing summary from the Warramboo survey area. 43 Table 5.7: Results of Pseudoscorpiones molecular analysis and corresponding species determinations. 43 Table 5.8: Results of Schizomida analysis and corresponding species determinations. 46 Table 5.9: Results of Scolopendromorpha analysis and corresponding species determinations. 47 Table 5.10: Results of Polydesmida analysis and corresponding species determinations from the Warramboo survey area. 48 Table 5.11: Results of Diplura analysis and corresponding species determinations. 49 Table 5.12: Results of Coleoptera analysis and corresponding species determinations. 49 Table 5.13: Interspecific variation of Coleoptera from both phases of sampling, in relation to reference species. 50 Table 5.14: Results of Thysaura analysis and corresponding species determinations. 51 Table 5.15: Results of Symphyla analysis and corresponding species determinations. 51 Table 5.16: Results of Isopoda analysis and corresponding species determinations. 53 Table 5.17: Interspecific variation of Isopoda from both phases of sampling, in relation to reference species. 53 Table 6.1: Summary of species recorded from the Warramboo survey area, including their conservation status and distribution. (Coordinates in GDA94, Zone 50; A summary of the WAM SRE categories is located in Appendix 5). 56 Table 6.2: Overall habitat suitability of the study area 58

6 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figures Figure 3.1: Location of the Warramboo survey area and study area in relation to the existing Mesa A/Warramboo mining operations. 14 Figure 4.1: Climate and weather graph depicting long-term and monthly averages for the year preceding the final field mobilisation. 22 Figure 4.2: Location of drill holes sampled for troglofauna within the Warramboo survey area and previously sampled sites within the study area. 24 Figure 5.1: Drill holes sampled and surface geology of the Warramboo study area (see Table 5.1 for geology codes). 29 Figure 5.2: Conceptual cross section of the Robe Pisolite within the study area (Above) and from the Warramboo survey area (Below) 31 Figure 5.3: Modelled Robe Pisolite contours and thickness above water table within the Warramboo study (data supplied by Rio Tinto). 32 Figure 5.4: Prospective habitat within the Warramboo survey and study areas in relation to sampling locations. 34 Figure 5.5: Location of buffered PECs relevant to the current study area. 36 Figure 5.6: Summary of previous study locations in relation to the current scope. 39 Figure 5.7: Location of desktop review troglofauna sampling locations and records from the Warramboo study area. 42 Figure 5.8: Locations of specimen collections from the Warramboo troglofauna survey area during the current survey. 44 Figure 5.9: Bayesian analysis of CO1 haplotypes of the pseudoscorpion family Hyiidae collected from the Warramboo survey area. 45 Figure 5.10: Excerpt from Bayesian analysis of CO1 haplotypes of the pseudoscorpion family Chthoniidae collected from the survey area. 46 Figure 5.11: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Schizomida collected from the survey area. 47 Figure 5.12: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Scolopendromorpha collected from the survey area. 48 Figure 5.13: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Diplopoda collected from the Warramboo survey area. 49 Figure 5.14: Subsection of Helix produced Bayesian analysis of CO1 haplotypes of the order Coleoptera collected from the survey area. 50 Figure 5.15: Excerpt of Bayesian analysis of 12s haplotypes of the order Thysanura collected from the Warramboo survey area. 51 Figure 5.16: Excerpt of Bayesian analysis of CO1 haplotypes of the order Symphyla collected from the Warramboo survey area. 52 Figure 5.17: Excerpt of the Helix Bayesian analysis of CO1 haplotypes of the isopod family Armadillidae collected from the survey area. 52 Figure 6.1: Species (current and previous records) of conservation significance or potential conservation significance in relation to habitat suitability. 60

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 7 Warramboo Troglofauna Assessment

Plates Plate 5.1: Representative photos of drill log cores from Robe Pisolite geologies HTP from site DD14WMB0001 (A); and TPH, from site WARDC0024 (B). 30 Plate 5.2: Dorsal view of Schizomida species Paradraculoides sp. ‘SCH003’ (T138500, from drill hole MEARC3790), collected from the survey area. 47

8 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

1.0 Project Summary

In accordance with the Rio Tinto scope of work, Table 1.1 presents a summary of survey and project information.

Table 1.1: Summary of project specifications and limitations. Project Name Mesa A Hub: Warramboo Troglofauna Assessment Level of Survey Level 2 troglofauna survey Location Approximately 45 km west of Pannawonica township Survey Area Size 62.32 km2 Survey Timing Surveys completed between June – October 2015 Relevant Regulatory • EPA Statement of Environmental Principles, Factors and Objectives (EPA Guidance Documents and 2016a); Applicable Legislation • Environmental Factor Guideline – Subterranean Fauna (EPA 2016b); • Technical Guidance - Subterranean fauna survey (EPA 2016c); and • Technical Guidance - Sampling methods for subterranean fauna (EPA 2016d). Key Survey Limitations • Presence of bias in availability of drill holes, as sampling tends to not extend far beyond the footprint of the target ore body and is focussed on particular rock types. • Lack of adequate taxonomic framework and specialist expertise. • Higher than usual failure rates for the groups Diplura, Polydesmida and Isopoda during genetic analysis, due to tissue degradation and size of specimens. • Reconciling historic specimens with recent collections was dependent on the age of, and access to, historical specimens. Sequencing success deteriorates significantly with specimen age.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 9 Warramboo Troglofauna Assessment

This page intentionally blank

10 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

2.0 Executive Summary

Rio Tinto is evaluating the development of the Mesa A Hub, located within the western portion of the Robe Valley in the Pilbara Region, Western Australia. The Mesa A Hub encompasses multiple project areas, including the further development of the Warramboo deposit, which has the potential to impact subterranean fauna communities (both troglobitic and stygobitic fauna) that are known to be prevalent within the Robe Valley.

Rio Tinto commissioned Biota Environmental Sciences (Biota) to complete a level 2 troglobitic fauna assessment to inform the evaluation of these potential impacts, which comprised two phases of sampling as well as a desktop review collating previous records from the study area. For the purposes of this report the term ‘survey area’ refers to the Warramboo boundary inclusive of all current sample sites, whereas the ‘study area’ refers to the wider area encompassed by the desktop review (inclusive of the survey area). The study area was selected in liaison with Rio Tinto to encompass continuous submerged high prospectivity habitat extending from the survey area.

The objective of this report is to provide the necessary inputs to inform an environmental impact assessment (EIA) of proposed mining activities within the survey area.

The deposits of the Robe Valley are the remnants of palaeo channel deposits, generally forming mesa-like landforms. The adjacent or surrounding landforms have been removed by historical erosion. The Warramboo deposit is situated west of Mesa A and occurs below the ground level of the coastal plain.

The desktop review confirmed that one Priority Ecological Community (PEC) recognised by the Department of Parks and Wildlife is present within the study and survey area, and a second PEC was identified by the desktop review from the study area. These are, respectively: • Subterranean invertebrate community of pisolitic hills in the Pilbara; and • Subterranean invertebrate communities of mesas in the Robe Valley region.

For the purposes of this report, the geology of the study area was divided into high (Robe Pisolite over 5 m thickness), medium (Colluvium and Robe Pisolite <5 m thickness) and low (all remaining geological units) habitat prospectivity. Habitats of high prospectivity accounted for 37.3% of the study area by area, with medium accounting for an additional 33%. Approximately 80% of the study area therefore represents potential troglofauna habitat, with the main geological units of relevance having also been recognised by the EPA as potential troglofauna habitat during past assessments.

Sampling for troglofauna has previously been conducted within the study area as part of past surveys. Since 2004, 10 targeted troglobitic fauna surveys have been conducted. Western Australian Museum, Biota, NatureMap and Atlas of Living Australia databases were all queried as part of the desktop review to collate the existing records from these surveys.

The desktop review yielded a total of 168 troglofauna specimens from 46 taxa recorded between 2004 and 2014. Fourteen taxa were unable to be identified to species level, and as such are not considered within this assessment. Polyxenida sp. Indet. dominated the previously documented fauna assemblage, but this taxon has since been determined as Lophoturus madecassus and is not a Short-range Endemic (SRE).

Troglobitic fauna sampling was completed under “Licence to Take Fauna for Scientific Purposes” No. SF010296, and consistent with methodologies in EPA Guidance and guidelines. Sampling was completed using baited colonisation trap deployment, supplemented by drill hole scraping.

Two phases of troglofauna sampling were conducted within the study area between June and October 2015 with 33 drill holes sampled. In total, 128 troglofauna traps were installed, with the

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 11 Warramboo Troglofauna Assessment same sites utilised during both phases. Scraping was completed at eight drill holes prior to installation of Phase 1 troglofauna habitat traps.

While no major rainfall events occurred during the sampling periods, almost 250 mm of rain fell in the three months prior to sampling. Past observations suggest this approximate timing after significant rainfall creates suitable conditions in subterranean habitats for troglofauna sampling.

Collected specimens were separated out into morphotypes and preserved in 100% ethanol, which allows for both morphological and molecular analyses. The majority of collected specimens were sequenced for variation at the mitochondrial cytochrome oxidase subunit I gene (CO1), with Thysanura specimens sequenced using the mitochondrial gene 12S. The conservation significance of the species recorded was categorised as conservation significant species (those formally listed as specially protected under the Wildlife Conservation Act 1950 or identified as Priority taxa by the Department of Parks and Wildlife), SRE species, potential SRE species and non- SRE species.

A total of 43 specimens were collected from the Warramboo survey area during the current survey, comprising 32 from Phase 1 and 11 from Phase 2. All 43 specimens were sequenced, which determined that 18 species were present representing nine orders. Thirteen of these species are previously unrecorded and 10 from the current study were recorded from single drill holes only or were singleton records.

Based on the desktop review, three species of conservation significance were previously recorded from study area, Paradraculoides anachoretus (order Schizomida), Ideoblothrus linnaei and Ideoblothrus sp. ‘Mesa A’ (both of the order Pseudoscorpiones). Paradraculoides anachoretus is listed as Schedule 3 at State level, while both pseudoscorpion species are listed as Priority 1. Management and monitoring measures for impacts on these species arising from the existing Mesa A/Warramboo operations is covered under the approved Mesa A Troglofauna Management Plan.

On the basis of the consolidated data set (rationalising both the desktop review and recent sampling), thirteen species from the survey area were classified as being potential SRE species. Overall these species were poorly collected and are considered data deficient, with nine of the species representing singleton collections, and a further three species with more individuals but all collected from single site locations. Single site distribution locations are very unlikely to represent the true distribution of these species, however, and this result is likely an artefact of sampling effects and population density in the locality.

Within the study area 29.8 km2 represents confirmed troglofauna habitat, with over 57% of the confirmed habitat within that being of high prospectivity. Overall 87% of the records of conservation significant and potential SRE species were recorded from within areas of high habitat prospectivity, supporting the habitat categorisation.

While inferred high-suitability habitat also extends southwest of the current survey area, collection results appear to differ between the survey area and previous collections south of the survey area. No species of conservation significance, or of potential conservation significance as SREs, that have been recorded from within the survey area are currently known from the inferred continuation of the prospective habitat southwest of the survey area. One strongly troglobitic order, Schizomida, which is well represented in the survey area, is currently unrepresented from the wider study area. While there are no known faults or other structural features in the geology known to potentially isolate this south-western inferred portion of the study area, this may indicate a discontinuity in the habitat or change in extent of deeper and more complex subterranean habitats that may be utilised by more strongly obligate taxa. It is also possible that these patterns may be explained by the influence of ecological sampling effects on past data sets, particularly the effect of rainfall. Specifically-designed sampling within the wider Warramboo study area would be necessary to further inform if the differences in fauna occurrence are an artefact of sampling conditions or due to other factors such as changes in subterranean habitat.

12 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

3.0 Introduction

Rio Tinto is currently evaluating the development of a number of iron ore deposits within the Robe Valley, located in the Pilbara region of Western Australia. One such prospective development, the Mesa A Hub, is located within the western portion of the Robe Valley, approximately 45 km west of Pannawonica. The Mesa A Hub encompasses multiple project areas including Warramboo (Figure 3.1).

Mining of the Warramboo deposit is currently approved (EPA 2007a), with the Mesa A Hub proposal entailing both above and below water table expansion to the current above water table operations. This has the potential to impact subterranean fauna communities (both troglobitic and stygobitic fauna) that are known to be prevalent within the Robe Valley. To inform the assessment of these impacts, Rio Tinto wishes to increase knowledge of these communities by collating existing data, determining knowledge gaps and completing additional phases of sampling.

Rio Tinto commissioned Biota Environmental Sciences (Biota) to complete the required troglobitic fauna assessment, which comprised two phases of sampling as well as a desktop review collating previous records from the study area.

3.1 Study Objectives and Scope

The scope of this study was to undertake an assessment of troglobitic fauna and troglofauna habitat suitability within the Warramboo study area (approximately 323.5 km2). The key objectives of this study were as follows: 1. conduct a desktop review of relevant data available from the survey area and study areas (Section 5.2); 2. conduct a two-phase sampling programme to document the troglofauna assemblage within the study area (Section 5.3); and 3. place the recorded fauna into a regional context and discuss potential conservation significance (6.2).

The study was planned and implemented as far as practicable in accordance with the following: • EPA Statement of Environmental Principles, Factors and Objectives (EPA 2016a); • Environmental Factor Guideline – Subterranean Fauna (EPA 2016b); • Technical Guidance - Subterranean fauna survey (EPA 2016c); and • Technical Guidance - Sampling methods for subterranean fauna (EPA 2016d).

3.2 Purpose of this Report

The purpose of this report is to inform an environmental impact assessment (EIA) of proposed mining activities within the survey area as part of the Mesa A Hub proposal.

This report presents the findings of the desktop review, and documents the methodology, sampling effort and results of the two-phase troglobitic fauna sampling. Both the field sampling and report are subject to specific limitations that are discussed in Section 4.7.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 13 Warramboo Troglofauna Assessment

Figure 3.1: Location of the Warramboo survey area and study area in relation to the existing Mesa A/Warramboo mining operations.

14 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

3.3 Background on Subterranean Fauna

Until recently, Australia was thought to lack habitat suitable for subterranean fauna. However, recent research (mostly with an emphasis on impact assessment) has revealed Australia to be highly diverse in subterranean fauna, with numerous areas of interest throughout the continent (Guzik et al. 2010). In Western Australia these zones of subterranean biodiversity can be found at Cape Range, Barrow Island, the Pilbara bioregion, the Yilgarn and the Nullarbor (Humphreys 2001, Page et al. 2008, Guzik et al. 2010).

Relatively recent surveys in the Pilbara have collected subterranean fauna from a range of geological units such as pisolitic iron formations, channel iron deposits, unconsolidated alluvium and sedimentary basalt (Marmonier et al. 1993, Biota 2004, 2006, 2010, 2011a, 2013a). This indicates that the suitability of a formation as habitat for subterranean fauna is mostly a function of the availability of habitable space (Marmonier et al. 1993, Humphreys 1999, Biota 2006), rather than a specific geological unit.

Subterranean fauna habitats are characterised by shared physical parameters that include a lack of light, stable temperature, limited nutrient infiltration from surface environments and a constant humidity (Juberthie 2000, Romero 2009). These habitat characteristics have resulted in convergence in body morphology evolution amongst many subterranean fauna. Morphological characteristics common to most subterranean fauna include reduced or lack of pigmentation, reduced or lack of eyes, and elongate body morphology and appendages adapted for sensory movement (Culver and Pipan 2009, Romero 2009).

Subterranean fauna can be categorised into two distinct ecological groups based on habitat: troglofauna and stygofauna. Troglofauna are a suite of fauna that survive only in air-filled cavities and interstices between the ground surface and the water table. Stygofauna comprise aquatic taxa occurring in groundwater aquifers and subterranean water bodies. Troglobites and stygobites are obligatory subterranean habitat dwellers, and while they may occur close to surface environments, are strongly adapted to subterranean environments. This makes such fauna unable or highly unlikely to survive surface conditions. A range of similarly adapted fauna known as troglophiles, trogloxenes and edaphobites also occur; these fauna use subterranean habitats opportunistically but are able to survive outside these stable environments. Studies from the Pilbara bioregion have demonstrated that these suites of fauna are not similarly restricted in range and are therefore unlikely to be impacted by small-scale developments (Biota and Helix 2012, Helix 2012).

Troglofauna in semi-arid Australia are thought to be relictual rainforest fauna; fauna adapted to humid environments, which retreated underground to cave systems during the aridification of Australia (in the late Miocene; Humphreys 1993). This is inferred from affinities of the taxonomic groups represented amongst the troglofauna with other extant taxa in tropical climates. Some invertebrate groups with troglobitic representatives include the Arachnida (e.g. Schizomida, Pseudoscorpiones and Araneae), Chilopoda (e.g. Scolopendrida), Diplopoda (e.g. Polydesmida and Haplodesmida), and Insecta (e.g. Diplura, Thysanura, Coleoptera and Blattodea). A single troglobitic vertebrate species of blind snake (Anilios longissimus) is known from Australia, collected from Barrow Island and Cape Range (Aplin 1998, Humphreys et al. 2013).

Due to their dependence on constant humidity, the dispersal and distribution of troglobitic fauna species tends to remain limited to individual blocks of inter-connected habitat, leading to long periods of population isolation and speciation. As a result, troglobitic fauna are considered to be of conservation significance, given that species often display extreme short-range endemism and may therefore be affected by relatively small-scale developments such as mining and construction.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 15 Warramboo Troglofauna Assessment

3.4 Terminology

For the purpose of this report, the following terms are used as defined below: Study area – Refers to the overall area encompassing the deposit and the area included in the desktop review (Figure 3.1). Survey area – Refers to the area surveyed as part of the current Mesa A Hub Warramboo troglofauna assessment (Figure 3.1). Survey – Refers to the two phases of sampling for the current scope of work.

16 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

4.0 Methodology 4.1 Desktop Review and Database Searches

4.1.1 Review of Previous Relevant Studies A literature review was completed to identify relevant previous surveys from within the study area. This included searches of Biota’s library database, GoogleScholar1 and the EPA’s website, in addition to the identification of previous surveys via database specimen records (Section 4.1.2). Reports and data supplied by Rio Tinto were also collated with the results of the literature search and reviewed.

4.1.2 Database Searches

The following databases were searched to assist with compilation of a list of potential species for the study area: 1. NatureMap: a collaboration between the Department of Parks and Wildlife and the Western Australian Museum (WAM). This database represents the most comprehensive source of information on the distribution of Western Australia's fauna, comprising records from the Fauna Survey Returns database and WA Threatened Fauna Database (both maintained by the Department of Parks and Wildlife) and the WAM Specimen Database. 2. Atlas of Living Australia (ALA): a collaborative project between academic collecting institutions, private individual collectors and community groups. The atlas contains occurrence records, environmental data, images and the conservation status of species throughout Australia. 3. WAM‘s Arachnid, Myriapod and Crustacea database. 4. Biota Internal Database: This database includes all of the subterranean fauna data collected by Biota within Western Australia

Details of database searches conducted are summarised in Table 4.1.

Table 4.1: Summary and location of database searches completed for the current scope Database Date Search Target Bounding Coordinates Easting (m E) Northing (m N) Biota, NatureMap and ALA 09/01/17 Troglofauna (All orders) 375570 7598200 384140 7607700 WAM 11/11/15 Crustacea and Arachnida 393957 7607800 376398 7599100 TECs and PECs 09/01/17 TECs and PECs 333000 7548000 752000 7649000 Environmentally 24/11/16 Environmentally sensitive 333000 7548000 Sensitive areas areas 752000 7649000

4.2 Habitat Characterisation

The likely habitats for troglobitic fauna in the study area were initially characterised using a combination of regional information and site-specific geological data. Inputs considered in this analysis, where available, included:

1. the spatial extent of the study area in order to define the geological units that may be affected; 2. regional surface geology mapping;

1 https://scholar.google.co.uk

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 17 Warramboo Troglofauna Assessment

3. mapping of thickness above water table of prospective geological units; 4. hydrological and hydrogeological information; 5. stratigraphic logging, images and information from drillholes in the study area; 6. previous sampling effort in the locality and the results of published and unpublished studies on troglobitic fauna (Section 4.1); and 7. identification of rock types that have previously yielded troglobitic fauna records from the study area and wider locality (Section 4.1), including reviewing studies that have investigated the nature of troglofauna habitat specific to the Robe Valley (e.g. Biota and Blandford 2013).

The habitat units identified through this process were assigned a prospectivity to support troglofauna based on the above inputs, and then revisited on completion of the sampling component of this study, to further investigate if the spatial distribution of the fauna records provide support for the habitat model.

We have categorised the prospectivity of the geological units within the study area as Low, Moderate and High prospectivity for troglofauna, based on the following characteristics: A) Presence of mesocaverns, vugs and interstitial spaces. B) Known hydration, weathering or significant cavity zones. C) Presence of clay lenses or impeding layers to maintain stable humidity. D) The known occurrence of troglofaunal communities from equivalent rock types during past Pilbara surveys. E) Occurs above water table within the survey area.

Habitat prospectivity categories were then assigned as follows: Low – Suitable geology only occurs below water table in the survey area. Rock type may have B), C) and E) characteristics but locally lacking suitable habitat space. Troglofauna not known from previous studies sampling of the same geology. Medium – Suitable geology likely or known to occur above the water table in the survey area (E). Geology known to have interstices or vugs (A) and troglofauna have occasionally been detected in similar rock types previously (D). Geology may be subject to seasonal inundation (e.g. alluvium and colluvium). Where known, units of high prospectivity were categorised as medium if less than 5 m in thickness. High – Majority (four or five) categories confirmed for the geological unit, including E). Troglofauna routinely recorded from same rock type (D).

Habitat suitability was visualised by plotting troglofauna collections within the mapped prospectivity categories, using a 500 m buffer as a conservative extent for singleton records. This allows definition of areas of highly prospective habitat and confirmed troglofauna records from those without confirmed records but that are likely to be highly prospective based on geological knowledge. Species determined to be widespread were excluded from this analysis. All area within these buffer zones is confirmed habitat, while the area outside is inferred.

4.3 Sampling Methodology

Troglobitic fauna sampling was completed under “Licence to Take Fauna for Scientific Purposes” No. SF010296 issued to Jason Alexander (Appendix 1). Methodology and approach were consistent with those outlined in EPA Technical Guidance; Sampling Methods for Subterranean Fauna (EPA 2016e) and Technical Guidance; Subterranean Fauna Survey (EPA 2016c). Similar methodologies have been used in previous Robe Valley subterranean fauna assessments (Biota 2006, 2009a, 2011b).

18 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

4.3.1 Troglofauna Sampling Troglobitic fauna were sampled using two methods, deployment of baited colonisation traps and drill hole scraping.

4.3.1.1 Colonisation Trapping Custom-built litter colonisation traps were suspended at intervals within drill holes located within the study area. Traps were constructed from 60 mm internal diameter PVC irrigation pipe cut to a length of 180 mm. Each trap had a series of 20 mm holes drilled into the side, and traps remained open at the upper end. Up to three traps were installed such that they were in contact with the wall of the sampled drill hole, facilitating fauna entry into the trap. Number of traps installed varied depending on depth of the sites and presence and depth of groundwater.

Leaf litter was gathered locally from the ground surface in the study area, particularly from the bases of Acacia shrubs. The collected litter was soaked in water and irradiated in a microwave oven for two minutes on maximum power setting. The microwave acted to kill any invertebrates present and assisted in the breaking down of organic matter. Wet litter was added to the traps, which were kept in sealed containers until immediately prior to insertion into the drill hole to avoid desiccation of leaf litter. After the installation of traps, the opening of each drill hole was sealed to maintain humidity and to avoid entry of surface fauna.

Traps were recovered from each drill hole after seven weeks and stored in labelled zip lock bags to prevent desiccation of potential troglofauna specimens during transportation to Perth.

Fauna specimens were recovered from the traps using specially designed Tullgren funnel units. Leaf litter from each trap was placed in a sieve under an aluminium lamp containing a 25-watt globe. This created a temperature of approximately 30°C at the surface of the leaf litter. A funnel below the leaf litter collected the fauna as they fell through the sieve, directing them into an attached vial of 100% ethanol. Leaf litter was left in the Tullgren funnels for a period of 24 hours, or until dry, after which time the bulked invertebrate sample was removed.

4.3.1.2 Troglofauna Scraping Troglofauna were also sampled by using reinforced stygofauna haul nets. These nets were constructed from 70 µm plankton mesh and had 100 mm apertures attached to a weighted catch jar.

Troglofauna scraping was completed prior to the installation of troglofauna colonisation traps on the first phase. A net of suitable size was lowered to the bottom of each drill hole before being slowly hauled to the surface whilst scraping the wall of the drill hole, dislodging any fauna on the wall. Each drill hole was scraped a minimum of four times, to ensure the interior surface of the drill hole was adequately covered. On completion, fauna specimens were individually preserved in 100% ethanol.

The contents of the net, which included dry soil and root matter, were emptied into a container which was then filled with 100% ethanol to preserve any specimens that may have been in the soil and root matter for subsequent sorting in the field laboratory.

4.3.2 Data Management

Preliminary identification of subterranean fauna involved identification of specimens to order level, where possible, or separation of specimens into distinct morphotypes. Sorting was completed in Perth using dissecting microscopes (Olympus SZ40 and SZ61, magnification up to 40x). Morphotypes were then assigned a unique number based on drill hole name, date and method of collection. Specimens were preserved in 100% ethanol once separated out into morphotypes, which allows for both morphological and molecular analyses. Formal morphological taxonomic assessment by external experts was not completed as a part of this study and remains a limitation.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 19 Warramboo Troglofauna Assessment

4.4 Molecular Analysis

Molecular analysis of troglofauna groups was conducted to inform determinations of the number of species present, and compare the results with those obtained during previous surveys in the Robe Valley and wider Pilbara region. Yvette Hitchen of Helix Molecular Solutions (Helix) completed the molecular sequencing. Most specimens were sequenced for variation at the mitochondrial cytochrome oxidase subunit I gene (CO1; Appendix 5). Thysanura specimens were sequenced using mitochondrial gene 12S, as this gene has previously proved more effective at successfully yielding sequences from Pilbara Thysanura specimens, and therefore has superior reference data for comparison.

Dr Terrie Finston and Dr Oliver Berry (both of Helix) provided analysis and interpretation of the molecular data (Appendix 5). The molecular analysis used was a preliminary neighbour-joining approach using representative sequences from regional context data sets, followed by a phylogenetic analysis using the sequence data from this study (Appendix 5). This resulted in specimens from the current survey being placed into genetic lineages, which also included specimens from collections elsewhere in the region. Determination of putative species was then inferred from these genetic lineages based on the level of divergence between lineages of the same group (such as order or family), giving consideration to the relative variation within each lineage.

It should be noted that detailed morphological analysis was not completed for most specimens and the phylogenetic analysis was based on sequence data from a single gene. Therefore, all species arrived at by this approach should be considered putative unless previously described and fully determined by taxonomic specialists. Lineages with a divergence of less than 4.0% were considered a single species, whereas lineages with a divergence greater than 6.0% were considered separate distinct species for the purpose of this report, unless advised otherwise by Helix based on other data for the relevant taxonomic group. Lineages with intermediate divergences ranging from 4.0–6.0% were resolved to a preliminary level for the purposes of this report, with further input from Helix, and consideration to collection proximity, habitat characteristics and geology.

4.5 Categories of Conservation Significance

For the purpose of this report, the conservation significance of the fauna collected during this study, or records collated during the desktop review, was categorised as per Table 4.2

Table 4.2: Conservation classification used within this report. Category Description Conservation Species listed as Priority, Schedule or Vulnerable at State or Federal levels Significant species Confirmed Short-range Species where sufficient taxonomic expertise is available, and with adequate endemic (SRE) representation in WAM collections or genetic databases, that are known to be Species * limited in distribution based on the geological characteristics Potential SRE species * Species where there is insufficient taxonomic knowledge or too limited a number of collections to determine SRE status. Habitat, morphology, molecular or taxonomic data deficient, but belonging to groups that may display short-range endemism. Widespread Well-collected species, that are typically taxonomically well resolved. Species are (not an SRE) species * not confined by geological barriers. * Category based on WAM SRE guidelines (WAM 2014)

20 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

4.6 Survey Design

4.6.1 Survey Timing and Personnel

After a site and ground-truthing reconnaissance field mobilisation in April 2015, two phases of troglofauna sampling were conducted at Warramboo between June and October 2015 (Table 4.3).

Table 4.3: Summary of field mobilisations completed for the Warramboo Troglofauna survey area. Dates Personnel Purpose Phase 23rd – 28th April, 2015 Jason Alexander, Field reconnaissance and ground- - Michael Delaney truthing 3rd – 7th June, 2015 Jason Alexander, Troglofauna scraping, installation of 1 Penny Brooshooft troglofauna traps 6th – 8th August, 2015 Jason Alexander, Troglofauna trap recovery 1 Michael Delaney Installation of troglofauna traps 2 29th September – 2nd Jason Alexander, Troglofauna trap recovery 2 October, 2015 Penny Brooshooft

This study was managed by Jason Alexander, with Garth Humphreys, of Biota, providing directional input. Preliminary sorting of collected subterranean fauna was completed by Jason Alexander, Jacinta King, Andrew Sheppard, Tim Sachse, Chris Cole, Penny Brooshooft, Jenna Hyatt and Danielle Rayner, all of Biota.

4.6.2 Weather

Temperature and rainfall data were obtained from Mesa J mining operations approximately 42km east of the survey area. Long-term climatological reference data (rainfall data from 1971 – 2014, temperature data from 1971 – 2005) were obtained from the Bureau of Meteorology (BOM) weather station at Pannawonica (station number 5069, approximately 51 km east of Warramboo.

Sampling occurred between June and September 2015. While no major rainfall events occurred during the sampling periods, almost 250 mm of rain fell in the three months prior to sampling. Past observations (e.g. Biota 2006) suggest this approximate timing after significant rainfall creates suitable conditions in subterranean habitats for troglofauna sampling. This is supported by collection numbers of target fauna in Phase 1 versus Phase 2 (see Section 5.3).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 21 Warramboo Troglofauna Assessment

2 3

Figure 4.1: Climate and weather graph depicting long-term and monthly averages for the year preceding the final field mobilisation. Long-term temperature data 1971-2005, rainfall data 1971-2014; arrows indicate field mobilisation timing: Arrow 1: Phase 1 installation and troglofauna scrape; Arrow 2: Phase 1 recovery and Phase 2 installation; Arrow 3: Phase 2 trap recovery.

4.6.3 Sampling Effort A total of 33 drill holes were sampled within the survey area, which involved the installation of 128 troglofauna traps (Table 4.4, Figure 4.2). Troglofauna sampling drill holes were selected from a list of available sites to provide spatial coverage of the target CID and surrounding geologies. The same drill holes were sampled during both phases. Scraping was completed at eight drill holes prior to installation of troglofauna habitat traps.

Table 4.4: Name, location and sampling information of drill holes sampled for troglofauna within the Warramboo survey area. Drill hole Easting Northing Traps Installed Troglofauna Name (m E) (m N) Phase 1 Phase 2 Scrape MEARC2401 376945 7605874 2 2 MEARC3500 378199 7605666 2 2 MEARC3790 379796 7605620 2 2 Yes MEARC3811 379911 7605220 1 1 MEARC3814 379907 7605517 1 1 Yes MEARC4259 382940 7602963 3 2 MEARC4273 382991 7602721 2 2 MEARC4383 381850 7602435 3 2 MEARC4400 379855 7602466 1 1 MEARC4795 376177 7604750 2 2 MEARC4802 376200 7604053 1 1 Yes MEARC4923 379140 7606042 2 2 Yes MEARC4958 379603 7607061 2 2 MEARC4969 379894 7607277 2 2 Yes MEARC5015 382457 7602157 2 2

22 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Drill hole Easting Northing Traps Installed Troglofauna Name (m E) (m N) Phase 1 Phase 2 Scrape MEARC5017 382839 7602671 3 2 MEARC5038 381871 7601743 3 2 MEARC5053 381294 7601122 3 2 MEARC5069 380694 7601100 3 2 Yes MEARC5078 381271 7601911 2 2 Yes MEARC5091 381653 7602525 3 2 MEARC5093 381642 7602922 3 2 Yes MEARC5098 381446 7602720 1 1 MEARCUNK01 377526 7604742 2 2 RC12TOB0026 378980 7599942 3 2 RC13MEA0279 376781 7605110 1 1 RC13TOB0013 379157 7599709 2 2 TOBRC0009 380913 7600605 3 2 TOBRC0020 379527 7599960 2 2 TOBRC0023 379345 7599952 1 1 TOBRC0027 379515 7600362 3 2 TOBRC0028 379730 7600160 3 2 Total number of traps installed 70 58 - Total number of drill holes sampled 33 33 8 Coordinates in zone 50, datum GDA94

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 23 Warramboo Troglofauna Assessment

Figure 4.2: Location of drill holes sampled for troglofauna within the Warramboo survey area and previously sampled sites within the study area.

24 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

4.7 Limitations

Several limitations apply to this study, some of which are common functions of working on subterranean fauna, rather than functions of this specific study. These limitations include: 1. Sampling for troglofauna in the Robe Valley and wider Pilbara region relies on the use of drill holes, which are almost always installed as part of exploration drilling and thus focus on geological units of economic importance. This creates a bias in availability of drill holes for troglofauna sampling, as sampling tends to not extend far beyond the footprint of the target ore body and is focussed on particular rock types. This places limitations on determining wider species distributions and whether there are potential barriers to dispersal. This bias is extended to mapping contained within this report, such as habitat suitability (Section 6.3). 2. For several of the faunal groups there is a lack of adequate taxonomic framework and specialist expertise, both within Australia and internationally. Using genetic analysis in place of morphological identification for the majority of specimens, somewhat alleviates this limitation in this study, however the usefulness of genetic analysis is in turn limited by the number of reference specimens available for some groups (especially the Chilopoda and Symphyla) and that a single gene only has been sequenced here (Section 4.4). 3. DNA extraction and amplification for genetic analysis for the groups Diplura, Polydesmida and Isopoda had higher than usual failure rates due to degradation of the genetic material and insufficient genetic material. Despite multiple attempts, no sequence was recorded from 16 specimens from these three groups (Section 5.3.1). 4. Our ability to reconcile historical specimens from the desktop review with specimens collected during this survey was dependent on the age of, and access to, historical specimens. Sequencing success deteriorates significantly with specimen age and the available existing specimens were often unable to be sequenced as reference specimens (see point 3 above). 5. Conservation significance could not be assigned to taxa that could not be identified to species level (such as Diplura sp. Indet. or Trinemura sp. Indet.) as these specimens may corresponds to taxa already known from elsewhere or may represent species complexes. 6. Habitat suitability was plotted using all conservation significant and potential SRE species. While many of these species are currently classified as such due to singleton records, additional taxonomic work may determine they have widespread distributions. This inclusion may skew habitat suitability and result in greater numbers and potential collections from low habitat prospectivity.

Despite these limitations, the study provides an assessment that meets current EPA guidance (Section 3.1) and is adequate to inform the forthcoming EIA for the Mesa A Hub proposal.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 25 Warramboo Troglofauna Assessment

This page intentionally blank.

26 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

5.0 Results 5.1 Study Area Habitat Characterisation

Core habitat for subterranean fauna is primarily a function of available space, ability to maintain a constantly high humidity (for troglofauna) and the potential for nutrient input from surface systems (Humphreys 1991, Wilkens et al. 2000, Biota and DC Blandford & Associates 2013).

The following sections provide an initial description of the physical aspects relevant to the subterranean fauna communities pertinent to the current scope. Section 6.3 revisits this preliminary habitat characterisation and relates this to records of troglobitic fauna obtained from this study.

5.1.1 Geology of the Study Area

The Channel Iron Deposits (CID) of the Robe Valley are the remnants of paleo channel deposits, generally forming variable mesa-like landforms. The adjacent landforms have been removed by historical erosion processes to expose the CID rock as the basal unit of the younger mesa landscape (DC Blandford & Associates 2009). Robe Pisolite (which comprises the CID) forms the erosion-resistant upper surface and structural benches of the mesas (Hocking and Van de Graaff 1987). The current Warramboo deposit is situated west of Mesa A. The current proposed extension is superficially an extension of both existing Warramboo and Mesa A deposits. The Warramboo CID predominantly dips below the coastal plain, representing the western end of the cuesta landform that includes Mesa A.

The geology at Warramboo is simple with no complex structures, faults or fractures. Warramboo deposit is channel fill within the paleo channel of the Robe River, which is incised into Cretaceous age rocks in the local area. The Robe Pisolite exists as thin sheets filling the paleo channel where erosion has not reached the total depth of the original river channel. Warramboo covers parts of this paleo channel, which is the continuation of the Robe Pisolite deposit in Mesa A. These tenements are located downstream from the currently exposed mesa deposits of the main Robe River paleo channel.

The Warramboo study area comprises 11 surface geological units, with colluvium and lancustrine deposits comprising over 50% of the surface geology (Table 5.1, Figure 5.1). Robe Pisolite comprises 15.9% of the study area surface geology.

Table 5.1: Surface geology of the Warramboo troglofauna study area and potential for occurrence of subterranean fauna. Age Code Description Hectares Habitat Suitability (percentage of Characteristics study area) (Section 4.2) Cainozoic Qg Colluvium: Unconsolidated to loosely 6,309.6 A, D, E consolidated slope deposits. (19.5%) Qi Lacustrine deposits: clay, silt; saline in 10,109.0 C, E part, flood deposits. (31.3%) Qp Eluvium: clay and sandy clay plain with 4,495.0 C, E gilgais; intermittent veneer of alluvium; (13.9%) residual deposits of sand, gravel, and pebbles. Unit predominantly above water table. Qpt Eluvium: lightly cemented, low slope 1,299.2 E deposits; shale and limestone (4.0%) fragments Tp Robe Pisolite: Pisolitic limonite deposits. 5,129.1 A, B, C, D, E Occurs along old river channels. (15.9%) Mesozoic Kn Conglomerate, arenite and siltstone 2,826.2 E with some fissile mudstone. (8.7%)

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 27 Warramboo Troglofauna Assessment

Age Code Description Hectares Habitat Suitability (percentage of Characteristics study area) (Section 4.2) Kny Yarraloola Conglomerate: Poorly sorted 769.8 A, C, E conglomerate with shale, claystone (2.4%) lenses and interbedded sandstone. Precambrian Wa Ashburton: wacke, mudstone, 532.6 E ferruginous mudstone interbedded with (1.6%) sandstone and dolomite intruded by doleritic sills Proterozoic Ma Warramboo Sandstone: Interbedded 812.0 E massive and flaggy quartz sandstone (2.5%) and shale Mk Kiangi Creek Formation: Quartz 18.3 E sandstone, siltstone, mudstone, (0.1%) dolomite, and minor conglomerate. Pg Granite Medium grained leucocratic 47.5 E granite (0.1%) Total 32,348.3

28 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figure 5.1: Drill holes sampled and surface geology of the Warramboo study area (see Table 5.1 for geology codes).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 29 Warramboo Troglofauna Assessment

The target pisolite deposit is predominantly a flat lying lenses of mineralisation, which can be broken down into sub-categories in profile. This profile through the deposit is described in detail from top to bottom (data provided by Rio Tinto; Table 5.1, Figure 5.2), below: • Hardcapped Tertiary Pisolite (HTP) - between 5 and 10 m thick and consists of massive and vitreous goethite. Contains many vugs and cavities (Plate 5.1). • Tertiary Pisolite Hard (TPH) - is usually found at the base of the HTP unit and is of variable thickness up to 40m. Pisoliths are of variable diameter with a core of hematite or goethite, goethite cortices and a matrix of goethite (Plate 5.1). • Tertiary Pisolite Mixed (TPM) - underlies TPH is a pervasively altered remnant of TPH. Alteration occurred as the reactive air/water interface moved through the Pisolite body during the process of topographic inversion. Cavities are more common than in TPH. • Tertiary Pisolite Clay (TPC) - bands of predominantly clay rich material mixed throughout tertiary pisolite. • Tertiary Pisolite Denatured (TPD) - Discrete, semi-continuous unit of enriched pisolite that ranges in thickness from 2 m to 8 m, but is typically 4 m or less. • Tertiary Pisolite Basal (TPB) - consists of coarse pisolite, between 7 and 12 mm diameter in a white clay matrix. The basal pisolite is not always present and is rarely seen in outcrops. While variable within this unit, locally TPB is unlikely to contain cavities.

Below this CID stratigraphy lays the basement lithologies, which consist mostly of submerged rocks of the Ashburton Formation and the Boolgeeda Iron Formation of the Hamersley Group.

B) Plate 5.1: Representative photos of drill log cores from Robe Pisolite geologies HTP from site DD14WMB0001 (A); and TPH, from site WARDC0024 (B). (Site locations displayed in Figure 5.1)

30 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figure 5.2: Conceptual cross section of the Robe Pisolite within the study area (Above) and from the Warramboo survey area (Below) (ALL – Represents alluvium; remaining codes as per Table 5.1) (cross section location displayed in Figure 5.1 and Figure 5.3).

The mapped CID robe pisolite extends southwest beyond the survey area before a sharp meander in the original river system in a north-westerly trend. The water table in that area lies variable between 9 – 24 m below ground level, meaning that at its thickest, this CID extension varyingly lays between 5 and 26 m thick above water table (Figure 5.3).

5.1.2 Habitat Prospectivity within the Study Area

The physical characteristics of geological units known to provide habitat for troglofauna include fractures, caverns, vugs, or interstices sufficient in size to accommodate troglobitic fauna (Section 4.2). Studies in the Pilbara region have previously found that geological units such as colluvium and pisolite (as occur at Warramboo; Table 5.1; Figure 5.1) are suitable troglofauna habitat, the latter particularly when weathered and hydrated (Biota 2006, 2014a, 2015a). In addition to habitat space for troglobites, these stratigraphic units are often associated with other lithologies with important hydrological functions for troglofauna habitat, such as impeding layers and clay lenses which store infiltrated water from recharge events, maintaining humidity in the system (Biota and DC Blandford & Associates 2013). The combination of these geological units provide for percolation of water and associated nutrients from surface habitats.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 31 Warramboo Troglofauna Assessment

Figure 5.3: Modelled Robe Pisolite contours and thickness above water table within the Warramboo study (data supplied by Rio Tinto).

32 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

From the habitat suitability characteristics outlined in Table 5.1, two geological units were identified as prospective habitats (High and Medium habitat prospectivity; Figure 5.4) and eight identified as non-prospective habitats (Low habitat prospectivity; Table 5.2).

Table 5.2: Prospectivity of the geological units within the study area to provide troglofauna habitat. Habitat Geological Unit Proportion of study Prospectivity surface area (%) High Robe Pisolite (Tp; >5m thickness (Figure 5.3)) 20.7 Medium Colluvium (Qg) 43.6 Robe Pisolite (TP; <5m thickness (Figure 5.3)) Low Lacustrine deposits (Qi), 35.7 Eluvium and Alluvium (Qp and Qpt), Conglomerate (Kn and Kny), Ashburton (Wa), Warramboo Sandstone (Ma), Kiangi Creek Formation (Mk), Granite (Pg) * Variable within this geology but low habitat prospectivity within the study area locality (data supplied by Rio Tinto)

The remaining geological units within the survey area are characterised by few vugs or cavities in which troglofauna can occur or occur predominantly below the water table (submerged geology is excluded as potential habitat for troglofauna). Limited sampling has been completed to date in Kny, Wa and Ma geologies. While these geological units may be utilised by troglofauna opportunistically, their physical characteristics suggest it its unlikely they provide important habitat for troglofauna communities.

The CID or Robe Pisolite geological unit is likely to be the primary habitat for troglobitic fauna, with potential for collection in adjacent colluvial units when in contact with the pisolite (Table 5.1). There are currently insufficient data and sampling effort from isolated colluvium deposits to determine if this geology provides a habitat without connection to core habitat units.

Both Robe Pisolite and colluvium geological units are identified by the EPA as potential troglofauna habitat (EPA 2016d), with the Robe Pisolite unit at Warramboo also listed as a Priority 1 PEC (“Subterranean invertebrate community of pisolitic hills in the Pilbara”) (Section 5.2.1).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 33 Warramboo Troglofauna Assessment

Figure 5.4: Prospective habitat within the Warramboo survey and study areas in relation to sampling locations.

34 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

5.2 Desktop Review Results

5.2.1 Priority Ecological Communities

Searches of the Department of Parks and Wildlife TEC, PEC and Environmentally Sensitive Areas database yielded two PECs relevant to the current study (Section 5.2.1). No environmentally sensitive areas were recorded from the search (Section 4.1.2).

One PEC is present within the survey area, and a second PEC was from the east of the study area. Both PECs are described in Table 5.3 and their extents shown in Figure 5.5.

Table 5.3: Description of PECs overlapping the study area. PEC Name Description Category Subterranean “Subterranean invertebrate communities of a series of isolated Priority 1 invertebrate pisolitic mesas in the Robe Valley near Pannawonica in the Shire of community of Ashburton in the State's Pilbara Region. Includes Mesa A, B, C, G pisolitic hills in the and K and mesas not yet surveyed. The mesas are remnants of old Pilbara valley infill deposits of the palaeo Robe River. Mesas are flat topped hills with a hard laterised goethite cap (Biota 2006). The cap is underlain by pisolite, which is made up of spherical accretions of iron minerals called pisoliths (the iron ore source), which often have small caverns and spaces between them. These interstices can be large enough to accommodate troglobitic fauna (Biota 2009b). It is assumed that the other ironstone hills also provide a similar habitat for troglobitic fauna that were located in these hills by Biota (Biota 2006).” Subterranean “Troglobitic faunal communities occur in extremely specialised Priority 1 invertebrate habitats and appear to require the particular structure and communities of hydrogeology associated with mesas (but also apparently with mesas in the Robe other ironstone hills in the Robe Valley Region; Biota 2006) to Valley region provide a suitable humid habitat (EPA 2007b). Specifically, the habitat is the humidified pisolitic strata (small round accretionary masses of rock). These troglobitic communities are believed to be relics of the late Miocene (23 to 5.3 million years BP), having arisen from tropical faunal lineages that descended into subterranean environments during the aridification of Australia (Biota 2006). Short range endemism is common in these fauna (Biota 2006). Eleven fauna taxa located in Mesa A, for example, was not located in any other mesa sampled. This high level of endemicity was the general pattern of distribution noted by Biota (Biota 2006) for mesas and other hills in the Robe Valley that were thought to be likely habitat for troglofauna.”

Both PECs are categorised as Priority One; Poorly-known ecological community. This category is defined as:

“Ecological communities that are known from very few occurrences with a very restricted distribution (generally ≤5 occurrences or a total area of ≤ 100ha). Occurrences are believed to be under threat either due to limited extent, or being on lands under immediate threat (e.g. within agricultural or pastoral lands, urban areas, active mineral leases) or for which current threats exist. May include communities with occurrences on protected lands. Communities may be included if they are comparatively well-known from one or more localities but do not meet adequacy of survey requirements, and/or are not well defined, and appear to be under immediate threat from known threatening processes across their range.”

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 35 Warramboo Troglofauna Assessment

Figure 5.5: Location of buffered PECs relevant to the current study area.

36 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

5.2.2 Previous Relevant Surveys Sampling for troglofauna has previously been conducted within the Warramboo survey area as part of other studies. These were completed between 2005 and 2014 and include the original Mesa A environmental impact assessment, Robe Valley mesas baseline troglofauna sampling, Mesa A operations compliance sampling and regional troglofauna studies (Biota 2005, 2006, 2011b, 2013b).

A desktop review was completed to identify relevant previous records from the survey area and the wider study area. Extensive subterranean fauna sampling has previously been conducted within the Robe Valley, including within the study area. Since 2004, 10 targeted troglobitic fauna surveys have been conducted overlapping the current study area (Appendix 6, Table 5.4, Figure 5.6).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 37 Warramboo Troglofauna Assessment

Table 5.4: Summary of previous surveys completed within the survey area and surrounds. Biota (2014b): Bennelongia MWH (2014): Mesa Single Phase Biota (2006): Mesa (2010): Robe Valley Biota (2012a): Mesa A Troglofauna Troglofauna A and Robe Valley Tenure: Congo Bore A Troglobitic Fauna Current Study Biennial Sampling for Tod Mesas Troglobitic and Dinner Camp Compliance Compliance Bore, Hubert Well, Fauna Survey Baseline Monitoring 2012 Monitoring: 2014 Congo Bore and Troglofauna Survey Highway Deposit Phase 1 2 6 * 1 1 1 1 Area Surveyed Warramboo Warramboo Mesa A, B, C, F, G, Dinner Camp, Mesa A and B Mesa A and B Tod Bore, Hubert H, K, 2402e, Todd Congo Bore Well, Congo Bore, Bore, Warramboo Highway Deposit Survey Timing 3 Jun' - 8 Aug' 8 Aug' - 2 21 Nov’ 2004 – 11 22 Sep' – 18 Nov' 23 May - 20 Jul' 25 Jun' - 5 Sep' 2014 16 Oct' - 8 Dec' 2015 Oct' 2015 Apr’ 2007 2010 2012 2013 (6 phases) Rain data (mm) Rain during Sampling 14.2 0 44 0 51.6 0 7.6 Rain 3 months preceding 254.8 28.4 627.4 26.8 47.4 110 1 Overall Sites Sampled Trapped 33 33 88 40 46 51 116 Number of traps 69 57 291 80 135 158 301 Scraped 8 0 0 40 0 50 65 Sites Overlapping current study area Trapped (# traps) 33 33 4 (11) 40 3 (11) 4 (11) 116 Scraped 8 0 0 40 0 4 65 Number orders collected 8 7 9 13** 6 7 6** overlapping current survey area • breakdown and comparison per phase in Appendix 6 ** some taxa regarded as troglobitic at the time of this study are considered to be edaphobitic

38 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figure 5.6: Summary of previous study locations in relation to the current scope.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 39 Warramboo Troglofauna Assessment

A summary of the troglobitic fauna records collated during the desktop review is provided in Table 5.5. Complete details of all previous records are displayed in Appendix 3. The database search bounding the Warramboo troglofauna study area yielded a total of 168 specimens, representing 46 taxa, from previous surveys. These specimens were recorded between 2005 and 2014, and identified using either morphological or molecular techniques (Appendix 3).

Table 5.5: Taxa and abundance (n) of troglofauna recorded from the desktop review. Taxonomy n Drill holes Order Family Species Araneae Oonopidae Oonopidae sp. 'AO014' 2 DCBRC_017, DCBRC_040 Prethopalpus sp. 'B21' 3 DCBRC_089, MEARC4191 Pholcidae Pholcidae sp. 'indet' ^ 1 A1536 Palpigradi Indeterminate Palpigradi sp. 'B6' 3 COBRC0022 Pseudoscorpiones Atemnidae Atemnidae sp. 'B2' 1 DCBRC_017 Chthoniidae Tyrannaochthonius sp. 1 MEADC2380 'Warramboo' * Tyrannochthonius basme 2 COBRC0010 Olpiidae Austrohorus sp. 'indet' ^ 1 COBRC0013, MEARC5053, MEARC5071 Olpiidae sp. 'indet' ^ 1 A1784 Olpiidae sp. 'POL013' 1 COBRC0013 Syarinidae Ideoblothrus linnaei * 1 MEA4316 Ideoblothrus sp. 'indet' ^ 1 COBRC0030 Ideoblothrus sp. 'Mesa A' * 1 MEA4063 Schizomida Hubbardiidae Paradraculoides 9 MEARC3092, MEADC2492, anachoretus * MEADC3188, MEARC2702, MEARC2740, MEARC3066, MEARC3073, MEARC4151 Schizomida Hubbardiidae Paradraculoides sp. 4 MEADC2381 ‘SCH003’ * Scolopendromorpha Cryptopidae Cryptopidae sp. 'CHI022' 1 COBRC0001 Cryptops sp. 'B14' 1 COBRC0019 Cryptops sp. 'indet' ^ 1 COBRC0001 Geophilomorpha Geophilomorpha sp. 6 COBRC0001, COBRC0008, 'indet' ^ COBRC0015 Indeterminate Scolopendrida sp. 'indet' ^ 1 MEARC3073 Polyxenida Lophoproctidae Lophoturus madecassus * 64 COBRC0022, MEARC2999, MEARC4318, MEARC4329, MEARC5038, MEARC5044, TOBRC0009 Diplura Indeterminate Diplura sp. 'indet' ^ 2 COBRC0022, MEARC2702 Japygidae Japygidae sp. 'B17' 2 COBRC0008 Parajapygidae Parajapygidae sp. 'B14' 1 COBRC0003 Projapygidae Projapygidae new genus 1 MEARC4318 sp. ‘nov’ * Projapygidae sp. 'B6' 4 DCBRC_001, TOBRC0043 Projapygidae sp. 'indet' ^ 2 COBRC0012 Coleoptera Curculionidae Curculionidae sp. 'B10' 2 COBRC0007 Indeterminate Coleoptera sp. 'indet' ^ 4 COBRC0001, COBRC0029, MEARC4329, MEARC5070 Zygentoma Indeterminate Thysanura sp. 'T001' 1 TOBRC0055 Nicoletiidae Atelurodes sp. 'B2' 12 DCBRC_017, DCBRC_089 Hemitrinemura sp. 'B4' 3 COBRC0007, COBRC0016 Trinemura sp. 'indet' ^ 3 MEARC5070, RC12COB0001, TOBRC0057

40 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Taxonomy n Drill holes Order Family Species Zygentoma Trinemura sp. 'B10' 10 MEARC4191 cont. Trinemura sp. 'B11' 1 COBRC0019 Trinemura sp. ‘Mesa A1’ 2 MEARC3098 Trinemura sp. ‘T1’ * 2 TOBRC0011 Cephalostigmata Indeterminate Symphyla sp. 'SYM025' 1 TOBRC0038 Scolopendrillidae Symphylella sp. 'B9' 4 COBRC0003, COBRC0009, COBRC0010, COBRC0030 Symphylella sp. 'indet' ^ 1 COBRC0015 Scutigerellidae Hanseniella sp. 'B10' 1 COBRC0033 Isopoda Armadillidae Armadillidae 2 MEARC5044, TOBRC0003 sp.’ISA009a/9b’ * Armadillo sp. 'B18' 5 DCBRC_0038, DCBRC_089, TOBRC0044 Armadillo sp. 'indet' ^ 1 DCBRC_0038 Indeterminate Isopoda sp. 'indet' ^ 4 COBRC0029, COBRC0035, MEARC4243, MEARC5044 Philosciidae Philosciidae sp. 'ISP051' 1 RC12COB0002 * Occurs within the survey area ^ Higher order resolution: not included within the assessment (Section 6.0)

Fourteen taxa were unable to be identified to species level, and as such are not considered further within this assessment (Table 5.5; Appendix 3). Four taxa belong to described species, Lophoturus madecassus, Tyrannochthonius basme, Ideoblothrus linnaei and Paradraculoides anachoretus.

Lophoturus madecassus was the highest abundance species recorded, comprising 36% of the total number of specimens. These specimens are not considered further in relation to the study area as previous studies have indicated that these Polyxenida represent a taxon that is widespread through the Pilbara (Biota and Helix 2012), and in fact has a circum-tropical global distribution (Megan Short, pers. comm. 2016). They are therefore not SRE taxa or of conservation significance.

Three species, Schizomida species Paradraculoides anachoretus, Pseudoscorpiones species Ideoblothrus linnaei and ideoblothrus sp. ‘Mesa A’, are listed as Schedule 3, P1 and P1 respectively in Western Australia (Section 6.2).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 41 Warramboo Troglofauna Assessment

Figure 5.7: Location of desktop review troglofauna sampling locations and records from the Warramboo study area.

42 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

5.3 Survey Results

A total of 43 troglomorphic specimens were collected from across the Warramboo survey area, comprising 32 from Phase 1 and 11 from Phase 2 (Table 5.6; Figure 5.8). The specimens represented two phyla, seven classes and 9 orders, and 17 taxa, comprising five indeterminate taxa and 12 species-level taxa (Section 5.3.1).

The orders Isopoda and Schizomida were the greatest contributors to faunal composition, comprising 30.4% and 23.9% of the total number of potential troglofauna specimens collected from the survey area.

All 43 specimens were morphologically identified to order level and subsequently sequenced (Section 4.4) to assign species-level designations (Table 5.6).

Table 5.6: Order-level identifications and sequencing summary from the Warramboo survey area. Taxonomy Number Collected Genes Sequenced Phylum Class Order (Common Name) Phase 1 Phase 2 12S CO1 Arthropoda Arachnida Pseudoscorpiones 2 1 - 3 (Pseudoscorpion) Schizomida (Schizomid) 8 2 - 10 Chilopoda Scolopendromorpha - 1 - 1 (Centipede) Diplopoda Polydesmida (Polydesmid 5 2 - 7 millipede) Diplura Diplura (Earwig) 1 1 - 2 Insecta Coleoptera (Beetle) 3 - - 3 Thysanura (Silverfish) 1 1 2 - Symphyla Symphyla (Symphyla) 1 - - 1 Crustacea Malacostraca Isopoda (Slater) 11 3 - 14 Total 32 11

A detailed account of each potential troglofauna order, including the results of genetic analysis, follows in Section 5.3.1.

In addition to the 43 potential troglofauna specimens, a large number of edaphobitic specimens were collected. As is usual for troglobitic fauna surveys in the Pilbara the majority of collected specimens were edaphobitic, or specimens derived from surface environments. A complete list of specimens recorded is appended (Appendix 2), however for the purpose of this assessment, these specimens are not discussed further.

5.3.1 Annotated Account of Collected Troglofauna

5.3.1.1 Pseudoscorpiones Three specimens from the order Pseudoscorpiones were successfully sequenced from the survey area (Table 5.7, Figure 5.8). Genetic analysis showed that the collected specimens represented two genetic lineages, which were equivalent to species, from two families, Hyiidae and Chthoniidae (Table 5.7, Figure 5.8, Figure 5.9).

Table 5.7: Results of Pseudoscorpiones molecular analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Chthoniidae Pseudoscorpiones PCH012 Chthoniidae sp. ‘PCH012’ 2 MEARC3814 Hyiidae Pseudoscorpiones PH006 Hyiidae sp. ’PH006’ 1 RC13MEA0279

The species collected from the family Chthoniidae, Chthoniidae sp. ’PCH012’, differed from closest reference specimens, Chthoniidae sp. ‘PCH050’, collected from nearby Mesa B, by 8.5% (Appendix 4).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 43 Warramboo Troglofauna Assessment

Figure 5.8: Locations of specimen collections from the Warramboo troglofauna survey area during the current survey.

44 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

The Hyiidae species collected from Warramboo, Hyiidae sp. ’PH006’, was most closely related (11.3% divergence) to a Buckland Hills species, collected approximately 79 km east-southeast of the current survey area.

The level of CO1 divergence displayed between Hyiidae sp. ‘PH006’, Chthoniidae sp. ‘PCH012’, and reference specimens representing previously identified species, is sufficient for these two lineages to be classed as discrete species based on molecular analysis (see Appendix 4). Both of these species are currently known from single locations and are poorly represented in collections (Section 6.2) and are considered new species.

Figure 5.9: Bayesian analysis of CO1 haplotypes of the pseudoscorpion family Hyiidae collected from the Warramboo survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured block indicates specimens belonging to a single species.)

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 45 Warramboo Troglofauna Assessment

Figure 5.10: Excerpt from Bayesian analysis of CO1 haplotypes of the pseudoscorpion family Chthoniidae collected from the survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured block indicates specimens belonging to a single species.)

5.3.1.2 Schizomida Ten specimens of the order Schizomida were collected and sequenced from the Warramboo survey area, belonging to two phylogenetic lineages (Table 5.8, Figure 5.8). Both lineages are from the family Hubbardiidae, and are highly divergent and as such represent separate species.

Table 5.8: Results of Schizomida analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Hubbardiidae Schizomida Paradraculoides sp. 9 MEARC3790, MEARC3811, SCH003 ‘SCH003’ MEARC4400, TOBRC0023 Schizomida Paradraculoides sp. 1 MEARC4273 SCH004 ’SCH004/004a’

The first species, Paradraculoides sp. ‘SCH003’ (Plate 5.2), has previously been recorded from one drill hole in the Warramboo survey area (Figure 5.7). This drill hole (MEADC2381; Figure 5.7) is located within the currently approved mine area. This species diverges from Paradraculoides anachoretus (Mesa A) and Paradraculoides bythius (Mesa B) by between 7.5% and 9.5% (Appendix 4). The known distribution of the species Paradraculoides sp. ‘SCH003’ extends over approximately 4.9 km2, within the Warramboo deposit.

Broader analyses showed that the second species, Paradraculoides sp. ’SCH004/004a’ (T138502), which was only recorded from one specimen, is very closely related to specimens collected during a concurrently run study at Mesa L, Mesa M and Mesa N in the East Deepdale area (approximately 45 km east of the Warramboo collection location). The specimen differs from Mesa L specimens by and average of 0.57%, with a range of 0.12 – 0.87% sequence divergence (Helix unpublished data, Appendix 4), and therefore is very likely to belong to the same species. This record is most likely an artefact of sample contamination, which is discussed further in Section 6.1.

46 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Plate 5.2: Dorsal view of Schizomida species Paradraculoides sp. ‘SCH003’ (T138500, from drill hole MEARC3790), collected from the survey area.

The Schizomida of the Robe Valley are considered entirely troglobitic with distributions linked mostly with single mesas (such as Paradraculoides anachoretus), or occasionally two proximal mesas (such as Paradraculoides bythius; Biota 2006, 2011b). Paradraculoides sp. ‘SCH003’ is likely to display similar life history and distribution characteristics to the known Schizomida species inhabiting the Robe Valley mesas (Section 6.2).

Figure 5.11: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Schizomida collected from the survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured block indicates specimens belonging to a single species.)

5.3.1.3 Scolopendromorpha A single specimen of the order Scolopendromorpha was collected from the Warramboo survey area (Table 5.9, Figure 5.8). This specimen, assigned to the lineage Chilopoda CHI002, belongs to the genus Cryptops (family Cryptopidae) and is the first record of this species. Its putative species, Cryptops sp.’CHI002’ differs from other reference species of the same family by between 18.0 and 28.1% sequence divergence (Appendix 4).

Table 5.9: Results of Scolopendromorpha analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Cryptopidae Chilopoda CHI002 Cryptops sp.’CHI002’ 1 MEARC4383

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 47 Warramboo Troglofauna Assessment

Figure 5.12: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Scolopendromorpha collected from the survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicates specimens belonging to a single species.)

Troglobitic specimens belonging to the genus Cryptops have previously been collected from the Robe Valley at Mesas A, B, G and K (Appendix 4; Biota 2006, 2007, 2012b). While reference data are somewhat limited, sequencing from previous studies and reference specimens indicate that Cryptops sp. ’CHI002’ is a distinct species previously not detected in the region.

5.3.1.4 Polydesmida Seven specimens of troglomorphic polydesmid millipede were collected from the Warramboo survey area (Table 5.10, Figure 5.8). Two specimens belong to the family Haplodesmidae, representing the species, Haplodesmidae sp. ‘DIHAP001’, were collected from a single drill hole in the survey area. The remaining five specimens failed to yield a viable sequence and therefore could not be determined to species level.

Table 5.10: Results of Polydesmida analysis and corresponding species determinations from the Warramboo survey area.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Indeterminate (DNA failed to amplify) Polydesmida sp. Indet ^ 5 MEARC2401, MEARC3500, RC13MEA0279 Haplodesmidae Haplodesmidae Haplodesmidae sp. 2 RC13MEA0279 DIHAP001 ‘DIHAP001’ ^ Higher order resolution: not included within the assessment further

Haplodesmidae sp. ‘DIHAP001’ has been recorded from multiple deposits within the Robe Valley, including Warramboo, Mesa B and Mesa C as well as from a location on the Hardey River (approximately 215 km southeast of Warramboo; Appendix 2, Figure 5.13). The intraspecific variation between populations between these deposits is 0.7%, indicating that gene flow is recent or ongoing across relatively broad spatial scales in these millipedes (Appendix 4). While troglobitic and SRE millipedes have previously been recorded from subterranean environments in the Pilbara (Humphreys et al. 2013), Haplodesmidae sp. ‘DIHAP001’ is unlikely to represent a troglobitic species as there is little evidence to indicate genetic subdivision or a restricted distribution based on geology and landforms. The known distribution of this species is 903 km2.

48 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figure 5.13: Excerpt of Helix produced Bayesian analysis of CO1 haplotypes of the order Diplopoda collected from the Warramboo survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicate specimens belonging to a single species.)

5.3.1.5 Diplura Two diplura specimens were collected from two drill holes, MEARC3792 and MEARC5017 (Table 5.11, Figure 5.8). However, DNA from both specimens failed to amplify and they were therefore unable to be sequenced or resolved to species level (Section 6.2).

Table 5.11: Results of Diplura analysis and corresponding species determinations. Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Indeterminate (DNA failed to amplify) Diplura sp. Indet ^ 2 MEARC3790, MEARC5017

^ Higher order resolution: not included within the assessment further

5.3.1.6 Coleoptera Genetic analysis of the three Coleoptera specimens collected identified three lineages, which correlated to three distinct species (Table 5.12, Figure 5.8).

Table 5.12: Results of Coleoptera analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Anthicidae Coleoptera CAN003 Stricticollis tobias 1 MEARC4400 Carabidae Coleoptera CCA001 Carabidae sp.’CCA001/012’ 1 MEARC3814 Curculionidae Coleoptera CCU005 Curculionidae 1 MEARC4400 sp.’CCU004/005’

Coleoptera lineage CAN003 (family Anthicidae) was assigned to an existing species, Stricticollis tobias, which has a global distribution (Telnov 2010). The Warramboo specimen differs from reference specimens used in the analysis (recorded outside of Australia) by 0.9% divergence (Section 6.2).

Lineage Coleoptera CCA001 from the survey area differed by 3.7% from a lineage recorded during a concurrent study at nearby Mesa B, suggesting that both belong to the same species (Figure 5.14; Appendix 4), Curculionidae sp. ’CCA001/012’. Collections of these two lineages is separated by 12.8 km.

This species differed from its most closely related reference species, Pterosticus mutus (GenBank reference species) by 12.4%, and from other specimens sequenced during this study by between 19.8% and 23.4% (Table 5.13). Due to this high level of divergence, this specimen represents a previously unrecorded species.

Coleoptera lineage CCU005 collected at Warramboo is very closely related (3.2 – 3.5% difference) to a lineage previously recorded from nearby Mesas A, B and C (Helix supplied data). These lineages represent a single species, Curculionidae sp. ’CCU004/005’ (Figure 5.14). This species differed from other reference taxa used in the analysis by between 20% and 25% sequence divergence. The known distribution for this species is 27.9 km2.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 49 Warramboo Troglofauna Assessment

Figure 5.14: Subsection of Helix produced Bayesian analysis of CO1 haplotypes of the order Coleoptera collected from the survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicate specimens belonging to a single species.)

Considering the low level of genetic diversity between specimens, and given that all species have known distributions from multiple, geomorphologically-separated deposits, it is likely that they represent troglophillic taxa (Section 6.2). There appears to be little geological restriction to dispersal for any of the collected Coleoptera species from the Warramboo study area.

Table 5.13: Interspecific variation of Coleoptera from both phases of sampling, in relation to reference species. Genetic divergence (%) Species Carabidae sp. Curculionidae sp. Stricticollis tobias ‘CCA001/012’ ’CCU004/005’ Stricticollis tobias Carabidae sp.’CCA001/012’ 19.8 Curculionidae sp. ’CCU004/005’ 23.7 23.4 Reference species Pterosticus mutus 21.0 12.4 24.9 Trigonopterus sp. 21.6 21.5 20.0

50 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

5.3.1.7 Thysanura Genetic analysis placed the two specimens of the insect order Thysanura collected from the current study area into two families, Nicoletiinae and Subnicoletiinae, and two species, Nicoletiinae sp. 'TN010' and Hemitrinemura sp. 'TS005’ (Table 5.14, Figure 5.8).

Table 5.14: Results of Thysaura analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Nicoletiinae Thysanura TN010 Nicoletiinae sp. 'TN010' 1 MEARC4383 Subnicoletiinae Thysanura TS005 Hemitrinemura sp. 'TS005’ 1 MEARC3814

Nicoletiinae sp. 'TN010' was recorded from drill hole MEARC4383 during the current study. Additionally, Nicoletiinae sp. 'TN010' has been collected from nearby Mesa B approximately 11.1 km east-northeast of the current survey area (Figure 5.15). This species differed from the closest reference specimen by 9.4%, which was collected during a previous study in the same survey area (Appendix 3).

Although closely related to a reference specimen from Christmas Creek (5.5% genetic divergence), Hemitrinemura sp. 'TS005’ is considered a distinct species. Given these specimens were sequenced with 12s, a slower evolving gene, this differentiation would equate to CO1 differentiation of over 7% (T. Finston, Helix pers. comm 2017). Hemitrinemura sp. 'TS005’ was recorded from a single drill hole within the survey area (Appendix 3).

Figure 5.15: Excerpt of Bayesian analysis of 12s haplotypes of the order Thysanura collected from the Warramboo survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicate specimens belonging to a single species.)

5.3.1.8 Symphyla A single Symphyla specimen was collected from the Waramboo troglofauna survey area (Figure 5.8). This specimen belongs to the previously unrecorded lineage Symphyla SYM026 from the family Scolopendrellidae, which differs from the closest available reference specimen by 16.7% (Appendix 4) and is therefore considered a new species, Scolopendrellidae sp. 'SYM026' (Table 5.15, Figure 5.17).

Table 5.15: Results of Symphyla analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Scolopendrellidae Symphyla SYM026 Scolopendrellidae sp. 'SYM026' 1 MEARC4273

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 51 Warramboo Troglofauna Assessment

Figure 5.16: Excerpt of Bayesian analysis of CO1 haplotypes of the order Symphyla collected from the Warramboo survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the survey area collected during the current study are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicate specimens belonging to a single species.)

5.3.1.9 Isopoda Fourteen isopod specimens were collected but nine were unable to be successfully sequenced and therefore could not be assigned to a lineage or species level taxon (Table 5.16). The remaining five specimens represented four lineages and three species all from the family Armidillidae (Table 5.16, Figure 5.8). Isopoda lineages 9a and 9b differed from each other by 3.8% and represent the same species based on their location and relatively low genetic divergence (Appendix 4).

One of the other lineages corresponded to a previously sequenced species (Armadillidae sp.’ISA009a/9b’), collected from the adjacent Mesa A deposit. The known distribution for this species is 8.7 km2. The remaining lineages correspond to species that were previously unrecorded (Figure 5.17).

Figure 5.17: Excerpt of the Helix Bayesian analysis of CO1 haplotypes of the isopod family Armadillidae collected from the survey area. (Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Warramboo specimens are displayed in pink. Reference and Genbank specimens displayed in black. Coloured blocks indicate specimens belonging to a single species.)

52 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Table 5.16: Results of Isopoda analysis and corresponding species determinations.

Taxonomy Number Drill Hole Family Genetic Lineage Species Name Recorded Indeterminate (failed to sequence) Isopoda sp. Indet ^ 9 MEARC3811, MEARC4795, TOBRC0020 Armadillidae Isopoda lineage ISA006 Armadillidae sp. ’ISA006’ 1 TOBRC0020 Isopoda lineage ISA007 Armadillidae sp. ’ISA007’ 1 TOBRC0020 Isopoda lineage ISA009a Armadillidae sp. 1 MEARC4923 Isopoda lineage ISA009b ’ISA009a/9b’ 2 MEARC2401, MEARC5053 ^ Higher order resolution: not included within the assessment further

Species Armadillidae sp. ’ISA006’ and Armadillidae sp. ’ISA007’ were more genetically similar to each other (6.7% divergence) than to Armadillidae sp. ’ISA009a/9b’ (20.7%). The species were highly divergent from the closest reference specimens, ranging from 19.7% to 24.2% sequence divergence (Table 5.17).

Table 5.17: Interspecific variation of Isopoda from both phases of sampling, in relation to reference species. Genetic Divergence (%) Species Armadillidae sp. Armadillidae sp. Armadillidae sp. ’ISA009a/9b’ ’ISA006’ ’ISA007’ Armadillidae sp. ’ISA009a/9b’ Armadillidae sp. ’ISA006’ 20.7 Armadillidae sp. ’ISA007’ 20.7 6.7 Reference Species Isopoda sp. ’mesa L’ 23.0 24.2 23.8 Troglarmadillo sp.’ISO005’ 21.9 22.7 22.7 Troglarmadillo sp.’ISA0021’ 21.6 19.7 20.0 Previous records data supplied by Helix.

While epigean isopods are routinely collected from leaf litter, and are less prone to extreme short- range endemism, recent genetic analysis of subterranean specimens have indicated that troglomorphic isopods can display habitat restriction (Biota 2015b, Helix 2015a, 2015b).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 53 Warramboo Troglofauna Assessment

This page intentionally blank.

54 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

6.0 Discussion 6.1 Recorded Troglobites Excluded from Assessment

One species of Schizomida documented by this study, is not discussed further in the conclusions of this report. Paradraculoides sp. ’SCH004/004a’, recorded from site MEARC4273 (Section 5.3.1.2), is the same species collected from Mesa L, M and N, approximately 45 km east of the Warramboo collection sites. Given the extreme short range endemism prevalent within the Robe Valley troglobitic fauna communities (Biota 2006, 2011b), and the Schizomida in particular, along with the low level of genetic divergence between the Warramboo specimen and the Mesa L species over such a large distance, it is probable that this record is an artefact. This may be a result of cross-contamination between traps while in transit or during on-site storage (e.g. this specimen moved between the stored traps through an accidental tear in the ziplock bags during transit). To provide context, specimens of the same species from Mesa N differ from Mesa L specimens by an average of 1.9%, over an approximately 1.4 km distance, whereas the MEARC4273 specimen differed by just 0.57% over 45 km (Section 5.3.1.2). While this specimen is still reported for transparency, completeness and to maintain a conservative approach, it is not considered to represent a species endemic to the Warramboo survey area and thereby not considered in Section 6.2.

6.2 Conservation Significance

A total of 22 troglofauna species are now known from the survey area, three of which have been identified as being of conservation significance, Paradraculoides anachoretus (order Schizomida), Ideoblothrus linnaei and Ideoblothrus sp. ‘Mesa A’ (order Pseudoscorpiones; Table 6.1). All three species were recorded during previous surveys and documented during the desktop review as occurring within the survey area. Paradraculoides anachoretus is listed as Schedule 3, while both pseudoscorpion species are listed as Priority 1 (Table 6.1). These species are currently known only from Mesa A, which is adjacent to and partially overlaps the current survey area.

No confirmed SRE species were recorded from the current study (Table 6.1). In total, 13 species from the survey area were classified as being potential SRE species. Overall these species were poorly collected and data deficient, with nine of the species representing singleton collections, and a further three species collected from single site locations (Table 6.1). This single site location is unlikely to represent the true distribution of these species however and is likely an artefact of sampling effects.

Currently these species have the following attributes of relevance to the assessment of conservation significance of troglobitic fauna: • species with very short range distributions based on available data; each species currently appears, to be restricted to either a local paleo channel system or an individual mesa; • relictual fauna representative of very old lineages; the lineages from which the contemporary troglofauna have been present in subterranean habitats since the late Miocene (at least the last 10 million years); and • similar distributions to species which are currently listed as conservation significant (Vulnerable, Schedule or Priority fauna) under State legislation and occur in similar habitats in the bioregion.

It is likely that the troglobitic species from the Warramboo survey area would be considered data deficient (requiring additional specimens and improvements in overall taxonomic frameworks), or potentially assigned a similar conservation status to the previously described troglobitic species endemic to the Robe Valley mesas (such as Paradraculoides anachoretus or Ideoblothrus linnaei). These potential SRE species are treated here as troglobitic and SRE fauna for the purposes of conservative environmental impact assessment.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 55 Warramboo Troglofauna Assessment

The remaining six species are considered Widespread from the survey area, which included Stricticollis tobias, a cosmopolitan species described from specimens outside Australia (Telnov 2010). The remaining five species, while varying between fair and poorly collected from an area less than 10,000 km2, are considered widespread based on habitat indicators and distributions known for other related taxa. These species were recorded from the current study and at other deposits around the Mesa A Hub and elsewhere in the Pilbara (Table 6.1). These specimens show little geological restriction and are likely to be troglophilic species.

56 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Table 6.1: Summary of species recorded from the Warramboo survey area, including their conservation status and distribution. (Coordinates in GDA94, Zone 50; A summary of the WAM SRE categories is located in Appendix 5). Species Name Distribution Collection New Singleton SRE Notes 2 (km ) Representation* Species Record ∞ Sub-Categories ^ Conservation Significant Species Ideoblothrus linnaei 0.4 Poor - - - Conservation significant, Listed as Priority 1 in WA. Three specimens known from Mesa A. Assessed as part of the Mesa A/Warramboo proposal (MS 756). Ideoblothrus sp. ‘Mesa A’ 1.3 Poor - - - Conservation significant, Listed as Priority 1 in WA. Three specimens known from Mesa A. Assessed as part of the Mesa A/Warramboo proposal (MS 756). Paradraculoides anachoretus 5.7 Good - - - Conservation significant, Listed as Schedule 3 in WA. Over 80 specimens known from Mesa A distribution. Assessed as part of the Mesa A/Warramboo proposal (MS 756). Confirmed SRE Species (None recorded) ------Potential SRE Species Hyiidae sp. ’PH006’ - Poor X X A. Data Deficient - D. Molecular evidence Cryptops sp.’CHI002’ - Poor X X A. Data Deficient - D. Molecular evidence Hemitrinemura sp. 'TS005’ - Poor X X A. Data Deficient D. Molecular evidence Scolopendrellidae sp. 'SYM026' - Poor X X A. Data Deficient D. Molecular evidence Armadillidae sp. ’ISA006’ - Poor X X A. Data Deficient D. Molecular evidence Armadillidae sp. ’ISA007’ - Poor X X A. Data Deficient D. Molecular evidence Tyrannochthonius sp. 'warramboo' - Poor - X A. Data Deficient D. Molecular evidence Projapygidae new genus sp. ‘nov.’ - Poor - X A. Data Deficient C. Morphology indicator Thysanura sp. ‘T001’ - Poor X X A. Data Deficient D. Molecular evidence

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 57 Warramboo Troglofauna Assessment

Species Name Distribution Collection New Singleton SRE Notes 2 (km ) Representation* Species Record ∞ Sub-Categories ^ Chthoniidae sp. ‘PCH012’ - Poor X X A. Data Deficient Multiple specimens from single location. D. Molecular evidence Trinemura sp. ‘T1’ - Poor - X A. Data Deficient Multiple specimens from single location. Trinemura sp. ‘Mesa A1’ - Poor - X A. Data Deficient Multiple specimens from single location. Assessed as part of the Mesa A/Warramboo proposal (MS 756). Paradraculoides sp. ‘SCH003’ 1.0 Fair - - A. Data Deficient D. Molecular evidence Widespread Species Armadillidae sp. ’ISA009a/9b’ ° 8.7 Fair - - B. Habitat Indicators Also collected from Mesas A (MEARC2999). D. Molecular Evidence Likely Troglophilic.

Carabidae sp.’CCA001/012’ ° <0.1 Poor X - B. Habitat Indicators Collected also from Mesa B (RC15MEB0107). Likely Troglophilic.

Curculionidae sp. ’CCU004/005’ ° 27.9 Fair X - B. Habitat Indicators Also collected from Mesas A (MEARC2657), B (GR15MEB0003, RC14MEB0060, RC14MEB0068, RC15MEB0216) and C (GR15MEC0019, RC15MEC0192). Likely Troglophilic. Haplodesmidae sp. ‘DIHAP001’ ° 903.7 Fair - - B. Habitat Indicators Also collected from Mesa B (DD14MEB0005), C D. Molecular Evidence (RC15MEC0193) and Hardey River (215 km south east of Warramboo). Species Troglophilic. Nicoletiinae sp. 'TN010' ° - Poor X - B. Habitat Indicators Collected also from Mesa B (MEBRC0020). Likely Troglophilic. Stricticollis tobias Worldwide Good - - B. Habitat Indicators Introduced species * Poor: <5 specimens, Fair: 5 – 15, Good: >15. ∞ includes multiple specimens collected from single site. ^ A. Data Deficient, B. Habitat Indicators, C. Morphology indicators, D. Molecular Evidence, E. Research and Expertise (more detail in Appendix 5). ° Molecular and habitat data indicates species not restricted.

58 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

6.3 Habitat Suitability

In total, 67.0 km2 of the 323.5 km2 study area represents high prospectivity habitat, with 17 km2 (5.3% of total surface area) confirmed as habitat from sampling using a 500 m buffer around collections (Table 6.2, Figure 6.1).

Overall 87.2% of collections of conservation significant and potential SRE species (Table 6.1) were recorded from areas of High habitat prospectivity, supporting the habitat assessment. All orders recorded from the desktop review and current surveys were represented in the high habitat prospectivity, with the exception of Palpigradi.

A total of 141.1 km2 of habitat was categorised as Medium prospectivity with 4.5 km2 (3.2%) confirmed as habitat. Species recorded from Medium habitat prospectivity include from the group Schizomida (n=1), Pseudoscorpiones (n=1) and Thysanura (n=1).

A total of 8.3 km2 (7.7%) of Low prospectivity geology was categorised as confirmed habitat due to troglofauna collection from 11 sites south of the survey area. Species recorded from Low habitat prospectivity include the groups Palpigradi (n=1), Pseudoscorpiones (n=2), Diplura (n=2), Coleoptera (n=1), Thysanura (n=1), Symphyla (n=2) and Scolopendromorpha (n=1). These taxa, with the exception of the Cryptopidae sp. 'CHI022' (Scolopendromorpha) and Olpiidae sp. ‘POL013’ (Pseudoscorpiones), have all been identified solely on morphologically and mostly during previous surveys. Many of these taxonomic groups also have limited local taxonomic expertise and data for some suggest they may be less obligate than other troglofauna of the study area, such as the Schizomida and Pseudoscorpiones.

Table 6.2: Overall habitat suitability of the study area Habitat Prospectivity High Medium Low Confirmed Habitat area (km2) 17.0 4.5 8.3 (Troglofauna Recorded) Inferred Habitat area (km2) 50.0 136.6 107.0 (No Troglofauna Recorded)

A total of 186.6 km2 is categorised as inferred High (50.0 km2) and Medium (136.6 km2) prospectivity habitat within the study area (Table 6.2). While no fauna records exist from these areas, geology and other subterranean habitat information suggest that these areas are likely to provided habitat for troglofauna.

While inferred high-suitability habitat also extends southwest of the current survey area, collection results appear to differ between the survey area and previous collections south of the survey area. No species of conservation significance, or of potential conservation significance as SREs, that have been recorded from within the survey area are currently known from the inferred continuation of the prospective habitat southwest of the survey area. One strongly troglobitic order, the Schizomida, represented by two species within the survey area, is currently unrepresented from the wider study area. While there are no known faults or other structural features in the geology known to potentially isolate this south-western inferred portion of the study area, this finding may indicate a discontinuity in the habitat or change in extent of deeper and more complex subterranean habitats that may be utilised by more strongly obligate taxa. It is also possible that these patterns may be explained by the influence of ecological sampling effects on past data sets, particularly the effect of rainfall (Biota 2006, 2013c). Humphreys (1991) also observed that troglofauna populations in karst can expand rapidly after rainfall due to increased food and humidity.

Two sampling phases have been completed within the wider study area outside the survey area to date (Bennelongia 2010, MWH 2014). This previous sampling was completed during relatively dry periods in 2010 and 2013 (September – December), which may reduce the number of obligate subterranean fauna recorded and could also explain differences in the recorded fauna assemblages. However sampling from the Robe Valley mesas in 2010 over the same period (from September to November), still recorded 22 Schizomida from three of the mesas, as well as additional troglobitic taxa (Biota 2011b).

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 59 Warramboo Troglofauna Assessment

Specifically-designed sampling after significant rainfall within the wider Warramboo study area would be necessary to further inform if the observed differences in fauna assemblage are an artefact of sampling conditions or due to other factors such as changes in subterranean habitat.

60 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Figure 6.1: Species (current and previous records) of conservation significance or potential conservation significance in relation to habitat suitability.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 61 Warramboo Troglofauna Assessment

6.4 Conclusions

Three species of conservation significance, Paradraculoides anachoretus, Ideoblothrus linnaei and Ideoblothrus sp. ‘Mesa A’, were recorded from the survey area from previous surveys. These species are endemic to Mesa A, which partially overlaps the current study and survey areas. These species are unlikely to be impacted by the development at Warramboo as they are predominantly located outside the study area at Mesa A, and are covered under their approved management plan (Biota 2009b). No species formally identified as being of conservation significance has been recorded from only the Warramboo survey area.

Thirteen species from the survey area are categorised as potential SREs, of which all are data deficient (singleton records or poorly represented in collections). Over 87% of conservation significant and potential SRE species are recorded from high habitat prospectivity geologies. This geology, Robe Pisolite (Tp), unit is used by the Department of Parks and Wildlife as a surrogate to define the spatial boundaries of the Subterranean invertebrate community of pisolitic hills in the Pilbara PEC, as the Department “assumed that the other ironstone hills also provide a similar habitat for troglobitic fauna that were located in these hills”. In the current study, specimens of some taxonomic groups were also collected in low abundance from surrounding geological units.

While additional sequencing was utilised in an attempt to resolve taxonomic uncertainty for geographically restricted potential SRE species, the age of reference specimens curated in the Western Australian Museum reduced sequencing success. While these species remain unresolved, they are currently display similar life histories to known troglobitic species of conservation significance in the Robe Valley. Conservatively then, these species should be considered as potentially troglobitic for the purpose of the forthcoming EIA.

Overall, sampling successfully took place across 33 sites sampled on consecutive phases, which incorporated troglofauna scraping and installation of 128 troglofauna traps. Survey and desktop review analysis yielded 64 taxa in total. Twenty-two species level taxa are now known to occur within the survey area, of which three are of conservations significance, 13 are potential SRE species and six are widespread.

Geological modelling of prospective habitat showed over 87% of collections recorded from high prospectivity habitat, supporting the identification of subterranean habitat presented here. Overall, within the study area, there is modelled 67 km2 of High prospectivity habitat, with this comprising 17 km2 of confirmed and 50 km2 of inferred troglobitic fauna habitat.

62 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

7.0 Glossary

Edaphobite Deep soil inhabitant. Endemic Native to or confined to a certain region. EPA Environmental Protection Authority of Western Australia. Epigean Fauna from above the soil level. HTP Tertiary pisolite: Hydrated/hardcap pisolite geology. Interspecific Variation between species. variation Intraspecific Variation within species. variation Karst Soluble-rock landscape; terrain with distinctive hydrology and landforms arising from a combination of high rock solubility and well-developed secondary porosity. Mesocaverns Underground voids in the size range 0.1 – 20cm, especially in karst and volcanic substrates. Pisolite Rock composed of pisoliths. Short-Range A species that has a naturally small distribution and is often characterised Endemic (SRE) by having poor dispersal capabilities, confinement to disjunct habitats and low fecundity. WAM guidelines for determining SRE status in Appendix 5. Species Complex A group of closely related species currently placed within a single species name. Stygobite / Fauna inhabiting the various types of groundwater. Stygofauna TPB Tertiary pisolite: Basal, clay and pisolite. TPC Tertiary pisolite: Clay bands. TPD Tertiary pisolite: Denatured and friable ore. TPH Tertiary pisolite: Competent hard pisolitic ore. TPM Tertiary pisolite: Mix of TPC, TPH and TPD. Troglobite / Species that do not exist outside caves. They may, however, occur in the Troglofauna superficial underground compartment or in the upper hypogean zone. Troglophile / Species that utilise, but are not restricted to, subterranean environments. Troglophilic Able to use surface environments for dispersal.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 63 Warramboo Troglofauna Assessment

This page intentionally blank.

64 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

8.0 References

Aplin, K. P. (1998). Three new blindsnakes (Squamata: Typhlopidae) from northwestern Australia. Records of the Western Australian Museum 19:1–12.

Bennelongia (2010). Robe Valley Tenure; Congo Bore and Dinner Camp Baseline Troglofauna Survey. Unpublished report prepared for Rio Tinto, Bennelongia Environmental Consultants.

Biota (2004). Mesa A and Bungaroo Creek Exploration Areas Subterranean Fauna Survey. Unpublished report prepared for Robe River Iron Mining Company, August 2004, Biota Environmental Sciences, Western Australia.

Biota (2005). Interim Results from Mesa A Troglofauna Sampling. Unpublished interim letter report prepared for Rio Tinto, Biota Environmental Sciences, Western Australia.

Biota (2006). Mesa A and Robe Valley Mesas Troglobitic Fauna Survey. Unpublished report prepared for Robe River Iron Associates, March 2006, Biota Environmental Sciences, Western Australia.

Biota (2007). Mesa K Remnant Mining Project Troglobitic Fauna Survey. Unpublished report prepared for Pilbara Iron, June 2007, Biota Environmental Sciences, Western Australia.

Biota (2009a). Mesa G Troglofauna Survey 2009. Unpublished report prepared for Rio Tinto Iron Ore, December 2009, Biota Environmental Sciences, Western Australia.

Biota (2009b). Mesa A Troglofauna Management Plan. Unpublished report prepared for Rio Tinto Iron Ore, November 2009, Biota Environmental Sciences, Western Australia.

Biota (2010). Yandicoogina Subterranean Fauna Assessment Phases I - V. Unpublished report prepared for Rio Tinto Iron Ore, December 2010, Biota Environmental Sciences, Western Australia.

Biota (2011a). WPIOP Basalt Quarry Troglobitic Fauna Pilot Survey Results. Unpublished letter report prepared for API Management, 21 January 2011, Biota Environmental Sciences, Western Australia.

Biota (2011b). Robe Valley Mesas Troglobitic Fauna Survey 2010. Unpublished report prepared for Rio Tinto Iron Ore, March 2011, Biota Environmental Sciences, Western Australia.

Biota (2012a). Mesa A Troglobitic Fauna Compliance Monitoring 2012. Unpublished report for Rio Tinto, December 2012, Biota Environmental Sciences, Western Australia.

Biota (2012b). Mesa G Troglobitic Fauna Assessment Phases I-VI. Unpublished report prepared for Rio Tinto, March 2012, Biota Environmental Sciences, Western Australia.

Biota (2013a). Bungaroo Subterranean Fauna Collections Summary; Phases 1 - 11. Unpublished report prepared for Rio Tinto, March 2013, Biota Environmental Sciences, Western Australia.

Biota (2013b). Rio Tinto Regional Troglobitic Fauna Study. Unpublished report prepared for Rio Tinto, March 2013, Biota Environmental Sciences, Western Australia.

Biota (2013c). Koodaideri Troglobitic Fauna Assessment Phase VI Addendum Report. Unpublished report prepared for Rio Tinto, December 2013, Biota Environmental Sciences, Western Australia.

Biota (2014a). Koodaideri Iron Ore Project Troglobitic Fauna Habitat Assessment. Unpublished report prepared for Rio Tinto, February 2014, Biota Environmental Sciences, Western Australia.

Biota (2014b). Single Phase Troglofauna Sampling for Tod Bore, Hubert Well, Congo Bore and Highway Deposit. Unpublished report prepared for Rio Tinto, May 2014, Biota Environmental Sciences, Western Australia.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 65 Warramboo Troglofauna Assessment

Biota (2015a). Baby Hope Downs Troglofauna Survey Phase 2. Unpublished report prepared for Rio Tinto, July 2015, Biota Environmental Sciences, Western Australia.

Biota (2015b). Red Hill Creek and Stage 1 Extension Subterranean Fauna Assessment. Unpublished report prepared for API Management, November 2015, Biota Environmental Sciences, Western Australia.

Biota, and DC Blandford & Associates (2013). Robe Valley Troglofauna Habitat Characterisation and Reconstruction Review. Unpublished report prepared for Rio Tinto, June 2013, Biota Environmental Sciences and DC Blandford & Associates, Western Australia.

Biota, and Helix (2012). Polyxenid Millipede Regional Molecular Analysis. Unpublished report prepared for Rio Tinto Iron Ore, December 2012, Biota Environmental Sciences and Helix Molecular Solutions.

Culver, D. C., and T. Pipan (2009). The Biology of Caves and Other Subterranean Habitats. Oxford University Press.

DC Blandford & Associates (2009). West Pilbara Iron Ore Project: An Investigation into the geodiversity of palaeochannel systems. Unpublished report prepared for API Management, DC Blandford & Associates.

EPA (2007a). Statement No. 756: Mesa A / Warramboo Iron Ore Project. Published on 21st November 2007, Environmental Protection Authority, Western Australia.

EPA (2007b). EPA Guidance Statement No. 54a: Sampling Methods and Survey Considerations for Subterranean Fauna in Western Australia. Environmental Protection Authority, Western Australia.

EPA (2016a). Statement of Environmental Principles, Factors and Objectives. Environmental Protection Authority, Western Australia.

EPA (2016b). Environmental Factor Guideline: Subterranean Fauna. Environmental Protection Authority, Western Australia.

EPA (2016c). Technical Guidance: Subterranean Fauna Survey. Environmental Protection Authority, Western Australia.

EPA (2016d). Technical Guidance - Sampling methods for subterranean fauna . Environmental Protection Authority, Western Australia.

EPA (2016e). Technical Guidance: Sampling Methods for Subterranean Fauna. Environmental Protection Authority, Western Australia.

Guzik, M. T., A. D. Austin, S. J. B. Cooper, M. S. Harvey, W. F. Humphrey, T. Bradford, S. M. Eberhard, R. A. King, R. Leys, K. A. Muirhead, and M. Tomlinson (2010). Is the Australian subterranean fauna uniquely diverse? Invertebrate Systematics 24:407 – 418.

Helix (2012). Report on the molecular systematics of Polyxenida from the Pilbara - update. Unpublished report prepared for Biota Environmental Sciences, 20 December 2012, Helix Molecular Solutions, Western Australia.

Helix (2015a). Report on the molecular systematics of Coleoptera and Isopoda from Buckland Hills. Unpublished report prepared for Biota Environmental Sciences, 14 July 2015, Helix Molecular Solutions, Western Australia.

Helix (2015b). Report on the molecular systematics of the subfauna of API Red Hill. Unpublished report prepared for Biota Environmental Sciences, 29 October 2015, Helix Molecular Solutions, Western Australia.

Hocking, R. M., and W. J. E. V. Van de Graaff (1987). The Geology of the Carnarvon Basin, Western Australia.

66 Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx Warramboo Troglofauna Assessment

Humphreys, G., J. Alexander, M. S. Harvey, and W. F. Humphreys (2013). The subterranean fauna of Barrow Island, northwestern Australia: 10 years on. Records of the Western Australian Museum Supplement 83:145–158.

Humphreys, W. F. (1991). Experimental re-establishment of pulse-driven populations in a terrestrial troglobite community. Journal of Animal Ecology 60:609–623.

Humphreys, W. F. (1993). Cave fauna in semi-arid tropical Western Australia : a diverse relict wet- forest litter fauna. Memoires de Biospeologie 20:105–110.

Humphreys, W. F. (1999). Relict stygofaunas living in sea salt, karst and calcrete habitats in arid northwestern Australia contain many ancient lineages. Pages 219–227 in W. Ponder and D. Lumney, editors. The Other 99% -The Conservation and Biodiversity of invertebrates. Royal Zoological Society of New South Wales, Mosman.

Humphreys, W. F. (2001). The subterranean fauna of Barrow Island, northwestern Australia, and its environment. Memoires de Biospeologie (International Journal of Subterranean Biology) 28:107– 127.

Juberthie, C. (2000). Chapter 1: The Diversity of the Karstic and Pseudokarstic Hypogean Habitats in the World. Pages 17–40 in H. Wilkens, D. C. Culver, and W. F. Humphreys, editors. Ecosystems of the World 30: Subterranean Ecosystems. Elsevier Science, Amsterdam.

Marmonier, P., P. Vervier, J. Giber, and M. J. Dole-Olivier (1993). Biodiversity in ground waters. Trends in Ecology & Evolution 8:392–395.

MWH (2014). Mesa A Troglofauna Biennial Compliance Monitoring: 2014. Unpublished report prepared for Rio Tinto, MWH.

Page, T. J., W. F. Humphreys, and J. M. Hughes (2008). Shrimps Down Under: Evolutionary Relationships of Subterranean Crustaceans from Western Australia (Decapoda: Atyidae: Stygiocaris). PLoS One 3:e1618. doi: 10.1371/journal.pone.0001618.

Romero, A. (2009). Cave Biology: Life in Darkness. Cambridge University Press.

Telnov, D. (2010). Ant-like flower beetles (Coleoptera: Anthicidae) of the UK, Ireland and Channel Isles. British Journal of Entomology and Natural History 23:99–118.

Wilkens, H., D. C. Culver, and W. F. Humphreys (Eds.) (2000). Ecosystems of the World 30: Subterranean Ecosystems. Elsevier Science, Amsterdam.

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx 67

Warramboo Troglofauna Assessment

Appendix 1

Licence to take Fauna for Scientific Purposes (licence: SF010296)

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:1080 Ph1&2 Rev0_v3.docx

Warramboo Troglofauna Assessment

Appendix 2

Raw Sampling Data

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx

Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC2401 376945 7605874 P1 Acarina - - 20 - Morphological Surface fauna MEARC2401 376945 7605874 P1 Acarina - - 20 - Morphological Surface fauna MEARC2401 376945 7605874 P1 Acarina - - 20 - Morphological Surface fauna MEARC2401 376945 7605874 P2 Acarina - - 20 - Morphological Surface fauna MEARC2401 376945 7605874 P2 Acarina - - 20 - Morphological Surface fauna MEARC3500 378199 7605666 P2 Acarina - - 20 - Morphological Surface fauna MEARC3500 378199 7605666 P2 Acarina - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Acarina - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Acarina - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Acarina - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Acarina - - 20 - Morphological Surface fauna MEARC3811 379911 7605220 P1 Acarina - - 20 - Morphological Surface fauna MEARC3814 379907 7605517 P1 Acarina - - 30 - Morphological Surface fauna MEARC3814 379907 7605517 P2 Acarina - - 20 - Morphological Surface fauna MEARC4259 382940 7602963 P2 Acarina - - 20 - Morphological Surface fauna MEARC4259 382940 7602963 P2 Acarina - - 20 - Morphological Surface fauna MEARC4273 382991 7602721 P2 Acarina - - 20 - Morphological Surface fauna MEARC4273 382991 7602721 P2 Acarina - - 20 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Acarina - - 70 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Acarina - - 160 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Acarina - - 120 - Morphological Surface fauna MEARC4383 381850 7602435 P2 Acarina - - 20 - Morphological Surface fauna MEARC4383 381850 7602435 P2 Acarina - - 20 - Morphological Surface fauna MEARC4400 379855 7602466 P2 Acarina - - 20 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Acarina - - 30 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Acarina - - 200 - Morphological Surface fauna MEARC4795 376177 7604750 P2 Acarina - - 20 - Morphological Surface fauna MEARC4795 376177 7604750 P2 Acarina - - 20 - Morphological Surface fauna MEARC4923 379140 7606042 P2 Acarina - - 20 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC4923 379140 7606042 P2 Acarina - - 20 - Morphological Surface fauna MEARC4958 379603 7607061 P2 Acarina - - 20 - Morphological Surface fauna MEARC4958 379603 7607061 P2 Acarina - - 20 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Acarina - - 30 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Acarina - - 30 - Morphological Surface fauna MEARC4969 379894 7607277 P2 Acarina - - 19 - Morphological Surface fauna MEARC4969 379894 7607277 P2 Acarina - - 20 - Morphological Surface fauna MEARC5015 382457 7602157 P1 Acarina - - 50 - Morphological Surface fauna MEARC5015 382457 7602157 P1 Acarina - - 20 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Acarina - - 20 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Acarina - - 20 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Acarina - - 40 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Acarina - - 17 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Acarina - - 70 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Acarina - - 20 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Acarina - - 20 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Acarina - - 21 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Acarina - - 20 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Acarina - - 20 - Morphological Surface fauna MEARC5038 381871 7601743 P2 Acarina - - 20 - Morphological Surface fauna MEARC5038 381871 7601743 P2 Acarina - - 20 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Acarina - - 180 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Acarina - - 200 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Acarina - - 200 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Acarina - - 20 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Acarina - - 20 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Acarina - - 40 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Acarina - - 20 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Acarina - - 20 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC5078 381271 7601911 P1 Acarina - - 21 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Acarina - - 20 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Acarina - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Acarina - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Acarina - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Acarina - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Acarina - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Acarina - - 20 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Acarina - - 20 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Acarina - - 20 - Morphological Surface fauna MEARC5098 381446 7602720 P1 Acarina - - 15 - Morphological Surface fauna MEARC5098 381446 7602720 P2 Acarina - - 20 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P1 Acarina - - 20 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P2 Acarina - - 20 - Morphological Surface fauna

RC12TOB0026 378980 7599942 P2 Acarina - - 17 - Morphological Surface fauna

RC12TOB0026 378980 7599942 P2 Acarina - - 20 - Morphological Surface fauna

RC13MEA0279 376781 7605110 P1 Acarina - - 12 - Morphological Surface fauna

RC13MEA0279 376781 7605110 P2 Acarina - - 20 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P1 Acarina - - 20 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P2 Acarina - - 11 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P2 Acarina - - 20 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status TOBRC0009 380913 7600605 P1 Acarina - - 24 - Morphological Surface fauna TOBRC0009 380913 7600605 P1 Acarina - - 20 - Morphological Surface fauna TOBRC0009 380913 7600605 P1 Acarina - - 20 - Morphological Surface fauna TOBRC0009 380913 7600605 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0009 380913 7600605 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0023 379345 7599952 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0027 379515 7600362 P2 Acarina - - 7 - Morphological Surface fauna TOBRC0027 379515 7600362 P2 Acarina - - 8 - Morphological Surface fauna TOBRC0028 379730 7600160 P2 Acarina - - 20 - Morphological Surface fauna TOBRC0028 379730 7600160 P2 Acarina - - 20 - Morphological Surface fauna MEARC3811 379911 7605220 P1 Aranea - - 4 - Morphological Surface fauna MEARC3811 379911 7605220 P2 Aranea - - 3 - Morphological Surface fauna MEARC4273 382991 7602721 P1 Aranea - - 1 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Aranea - - 1 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Aranea - - 1 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Aranea - - 3 - Morphological Surface fauna MEARC4795 376177 7604750 P2 Aranea - - 1 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Aranea - - 2 - Morphological Surface fauna MEARC5078 381271 7601911 P1 Aranea - - 5 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Aranea - - 2 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Aranea - - 4 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Aranea - - 3 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Aranea - - 2 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Aranea - - 2 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Aranea - - 2 - Morphological Surface fauna MEARC5098 381446 7602720 P1 Aranea - - 1 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Blattodea - - 2 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC5069 380694 7601100 P1 Blattodea - - 1 - Morphological Surface fauna MEARC3500 378199 7605666 P1 Coleoptera - - 2 - Morphological Surface fauna MEARC3500 378199 7605666 P1 Coleoptera - - 5 - Morphological Surface fauna MEARC3500 378199 7605666 P2 Coleoptera - - 2 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Coleoptera - - 2 - Morphological Surface fauna MEARC4923 379140 7606042 P1 Coleoptera - - 1 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Coleoptera - - 1 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Coleoptera - - 2 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Coleoptera - - 4 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Coleoptera - - 1 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Coleoptera - - 1 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Coleoptera - - 1 - Morphological Surface fauna Widespread MEARC4400 379855 7602466 P1 Coleoptera Anthicidae Stricticollis tobias 1 90737 Molecular Species Carabidae sp. 90754 Widespread MEARC3814 379907 7605517 P1 Coleoptera Carabidae 1 Molecular 'CCA001/012' Species Curculionidae sp. Widespread MEARC4400 379855 7602466 P1 Coleoptera Curculionidae 1 90755 Molecular 'CCU004/005' Species MEARC2401 376945 7605874 P1 Collembola - - 20 - Morphological Surface fauna MEARC3500 378199 7605666 P2 Collembola - - 1 - Morphological Surface fauna MEARC3500 378199 7605666 P2 Collembola - - 4 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Collembola - - 25 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Collembola - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Collembola - - 20 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Collembola - - 20 - Morphological Surface fauna MEARC3811 379911 7605220 P1 Collembola - - 15 - Morphological Surface fauna MEARC3811 379911 7605220 P2 Collembola - - 20 - Morphological Surface fauna MEARC3814 379907 7605517 P1 Collembola - - 30 - Morphological Surface fauna MEARC3814 379907 7605517 P2 Collembola - - 20 - Morphological Surface fauna MEARC4259 382940 7602963 P2 Collembola - - 15 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC4259 382940 7602963 P2 Collembola - - 20 - Morphological Surface fauna MEARC4273 382991 7602721 P2 Collembola - - 1 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Collembola - - 2 - Morphological Surface fauna MEARC4383 381850 7602435 P1 Collembola - - 1 - Morphological Surface fauna MEARC4383 381850 7602435 P2 Collembola - - 1 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Collembola - - 200 - Morphological Surface fauna MEARC4795 376177 7604750 P1 Collembola - - 150 - Morphological Surface fauna MEARC4795 376177 7604750 P2 Collembola - - 2 - Morphological Surface fauna MEARC4795 376177 7604750 P2 Collembola - - 20 - Morphological Surface fauna MEARC4923 379140 7606042 P2 Collembola - - 3 - Morphological Surface fauna MEARC4958 379603 7607061 P2 Collembola - - 20 - Morphological Surface fauna MEARC4958 379603 7607061 P2 Collembola - - 20 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Collembola - - 25 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Collembola - - 4 - Morphological Surface fauna MEARC4969 379894 7607277 P2 Collembola - - 20 - Morphological Surface fauna MEARC4969 379894 7607277 P2 Collembola - - 20 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Collembola - - 2 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Collembola - - 15 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Collembola - - 11 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Collembola - - 2 - Morphological Surface fauna MEARC5017 382839 7602671 P1 Collembola - - 40 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Collembola - - 3 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Collembola - - 20 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Collembola - - 13 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Collembola - - 4 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Collembola - - 7 - Morphological Surface fauna MEARC5038 381871 7601743 P2 Collembola - - 3 - Morphological Surface fauna MEARC5038 381871 7601743 P2 Collembola - - 6 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Collembola - - 3 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC5053 381294 7601122 P1 Collembola - - 10 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Collembola - - 13 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Collembola - - 2 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Collembola - - 6 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Collembola - - 40 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Collembola - - 100 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Collembola - - 100 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Collembola - - 3 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Collembola - - 10 - Morphological Surface fauna MEARC5078 381271 7601911 P1 Collembola - - 2 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Collembola - - 3 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Collembola - - 6 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Collembola - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Collembola - - 3 - Morphological Surface fauna MEARC5091 381653 7602525 P1 Collembola - - 1 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Collembola - - 20 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Collembola - - 20 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Collembola - - 7 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Collembola - - 10 - Morphological Surface fauna MEARC5098 381446 7602720 P1 Collembola - - 70 - Morphological Surface fauna MEARC5098 381446 7602720 P2 Collembola - - 20 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P1 Collembola - - 8 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P1 Collembola - - 5 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P2 Collembola - - 6 - Morphological Surface fauna

MEARCUNK01 377526 7604742 P2 Collembola - - 20 - Morphological Surface fauna

RC13MEA0279 376781 7605110 P1 Collembola - - 2 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status

RC13MEA0279 376781 7605110 P2 Collembola - - 1 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P1 Collembola - - 3 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P2 Collembola - - 1 - Morphological Surface fauna

TOBRC0009 380913 7600605 P1 Collembola - - 1 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0023 379345 7599952 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0027 379515 7600362 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0027 379515 7600362 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0028 379730 7600160 P2 Collembola - - 20 - Morphological Surface fauna TOBRC0028 379730 7600160 P2 Collembola - - 20 - Morphological Surface fauna Unresolved (species level MEARC3790 379796 7605620 P1 Diplura Indeterminate Diplura sp. Indet 1 90759 Morphological resolution needed) Unresolved (species level MEARC5017 382839 7602671 P2 Diplura Indeterminate Diplura sp. Indet 1 90772 Morphological resolution needed) MEARC2401 376945 7605874 P1 Diptera - - 1 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Diptera - - 1 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Diptera - - 1 - Morphological Surface fauna MEARC3811 379911 7605220 P1 Diptera - - 1 - Morphological Surface fauna MEARC4383 381850 7602435 P2 Diptera - - 1 - Morphological Surface fauna MEARC4958 379603 7607061 P2 Diptera - - 1 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Diptera - - 1 - Morphological Surface fauna MEARC4969 379894 7607277 P1 Diptera - - 1 - Morphological Surface fauna MEARC5015 382457 7602157 P2 Diptera - - 1 - Morphological Surface fauna MEARC5017 382839 7602671 P2 Diptera - - 1 - Morphological Surface fauna

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status MEARC5038 381871 7601743 P2 Diptera - - 1 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Diptera - - 2 - Morphological Surface fauna MEARC5098 381446 7602720 P2 Diptera - - 1 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Diptera - - 1 - Morphological Surface fauna TOBRC0020 379527 7599960 P2 Diptera - - 1 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Hemiptera - - 1 - Morphological Surface fauna MEARC3790 379796 7605620 P2 Hemiptera - - 1 - Morphological Surface fauna MEARC4923 379140 7606042 P1 Hemiptera - - 1 - Morphological Surface fauna MEARC4923 379140 7606042 P2 Hemiptera - - 6 - Morphological Surface fauna MEARC4923 379140 7606042 P2 Hemiptera - - 10 - Morphological Surface fauna MEARC5038 381871 7601743 P2 Hemiptera - - 1 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Hemiptera - - 1 - Morphological Surface fauna MEARC5093 381642 7602922 P2 Hemiptera - - 2 - Morphological Surface fauna MEARC5098 381446 7602720 P2 Hemiptera - - 1 - Morphological Surface fauna

RC13TOB0013 379157 7599709 P2 Hemiptera - - 1 - Morphological Surface fauna

MEARC4273 382991 7602721 P2 Hymenoptera - - 1 - Morphological Surface fauna MEARC4923 379140 7606042 P2 Hymenoptera - - 1 - Morphological Surface fauna MEARC4969 379894 7607277 P2 Hymenoptera - - 1 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Hymenoptera - - 1 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Hymenoptera - - 2 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Hymenoptera - - 3 - Morphological Surface fauna MEARC5091 381653 7602525 P2 Hymenoptera - - 11 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Hymenotera - - 2 - Morphological Surface fauna Surface fauna MEARC3811 379911 7605220 P2 Isopoda - - 1 - Morphological /pigmented Surface fauna MEARC3814 379907 7605517 P2 Isopoda - - 1 - Morphological /pigmented Surface fauna MEARC4969 379894 7607277 P1 Isopoda - - 1 - Morphological /pigmented

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status Armadillidae sp. Potential SRE/ TOBRC0020 379527 7599960 P1 Isopoda Armadillidae 1 C60729 Molecular ’ISA006’ Singleton Armadillidae sp. Potential SRE/ TOBRC0020 379527 7599960 P2 Isopoda Armadillidae 1 C60733 Molecular ’ISA007’ Singleton Armadillidae sp. Widespread MEARC2401 376945 7605874 P1 Isopoda Armadillidae 1 C60723 Molecular ’ISA009a/009b’ Species Armadillidae sp. Widespread MEARC4923 379140 7606042 P2 Isopoda Armadillidae 1 C60732 Molecular ’ISA009a/009b’ Species Armadillidae sp. Widespread MEARC5053 381294 7601122 P1 Isopoda Armadillidae 1 C60727 Molecular ’ISA009a/009b’ Species Specimen failed MEARC3811 379911 7605220 P1 Isopoda Indeterminate Isopoda sp. Indet 1 C60724 Morphological to provide sequence Specimen failed MEARC4795 376177 7604750 P1 Isopoda Indeterminate Isopoda sp. Indet 6 C60726 Morphological to provide sequence Specimen failed MEARC4795 376177 7604750 P1 Isopoda Indeterminate Isopoda sp. Indet 1 C60725 Morphological to provide sequence Specimen failed TOBRC0020 379527 7599960 P2 Isopoda Indeterminate Isopoda sp. Indet 1 C60774 Morphological to provide sequence MEARC4273 382991 7602721 P2 Isoptera - - 5 - Morphological Surface fauna MEARC5078 381271 7601911 P2 Isoptera - - 1 - Morphological Surface fauna RC13TOB0013 379157 7599709 P1 Isoptera - - 1 - Morphological Surface fauna MEARC3790 379796 7605620 P1 Pauropoda - - 1 - Morphological Surface fauna MEARC4273 382991 7602721 P1 Pauropoda - - 1 - Morphological Surface fauna Haplodesmidae Widespread RC13MEA0279 376781 7605110 P1 Polydesmida Haplodesmidae 1 T139893 Molecular sp. 'DIHAP001' Species Haplodesmidae Widespread RC13MEA0279 376781 7605110 P1 Polydesmida Haplodesmidae 1 T139894 Molecular sp. 'DIHAP001' Species Specimen failed Polydesmida sp. MEARC2401 376945 7605874 P1 Polydesmida Indeterminate 1 T138443 Morphological to provide Indet sequence Polydesmida sp. Specimen failed MEARC3500 378199 7605666 P1 Polydesmida Indeterminate 1 T138527 Morphological Indet to provide

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status sequence Specimen failed Polydesmida sp. MEARC3500 378199 7605666 P2 Polydesmida Indeterminate 1 T138444 Morphological to provide Indet sequence Specimen failed Polydesmida sp. MEARC3500 378199 7605666 P2 Polydesmida Indeterminate 1 T138527 Morphological to provide Indet sequence Specimen failed Polydesmida sp. RC13MEA0279 376781 7605110 P1 Polydesmida Indeterminate 1 T138445 Morphological to provide Indet sequence MEARC2401 376945 7605874 P1 Polyxenida - - 3 - Morphological Surface fauna MEARC3814 379907 7605517 P1 Polyxenida - - 5 - Morphological Surface fauna MEARC3814 379907 7605517 P2 Polyxenida - - 3 - Morphological Surface fauna MEARC5038 381871 7601743 P1 Polyxenida - - 3 - Morphological Surface fauna MEARC5053 381294 7601122 P1 Polyxenida - - 1 - Morphological Surface fauna MEARC5053 381294 7601122 P2 Polyxenida - - 4 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Polyxenida - - 8 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Polyxenida - - 1 - Morphological Surface fauna MEARC5069 380694 7601100 P1 Polyxenida - - 1 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Polyxenida - - 1 - Morphological Surface fauna MEARC5069 380694 7601100 P2 Polyxenida - - 7 - Morphological Surface fauna Chthoniidae sp. Potential SRE/ MEARC3814 379907 7605517 P1 Pseudoscorpiones Chthoniidae 1 T138463 Molecular 'PCH012' Single site only Chthoniidae sp. Potential SRE/ MEARC3814 379907 7605517 P2 Pseudoscorpiones Chthoniidae 1 T138531 Molecular 'PCH012' Single site only Potential SRE/ RC13MEA0279 376781 7605110 P1 Pseudoscorpiones Hyiidae Hyiidae sp. 'PH006' 1 T138464 Molecular Singleton Paradraculoides MEARC3790 379796 7605620 P1 Schizomida Hubbardiidae 1 T138500 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides MEARC3790 379796 7605620 P1 Schizomida Hubbardiidae 1 T139910 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides MEARC3790 379796 7605620 P1 Schizomida Hubbardiidae 1 T139911 Molecular Potential SRE sp. ‘SCH003’ MEARC3790 379796 7605620 P1 Schizomida Hubbardiidae Paradraculoides 1 T139912 Molecular Potential SRE

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Easting Northing WAM Notes / Specimen Drill Hole Name Phase Order Family Species N ID Method (m E) (m N) Number Status sp. ‘SCH003’ Paradraculoides MEARC3790 379796 7605620 P1 Schizomida Hubbardiidae 1 T138499 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides MEARC3790 379796 7605620 P2 Schizomida Hubbardiidae 1 T138553 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides MEARC3811 379911 7605220 P1 Schizomida Hubbardiidae 1 T138501 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides MEARC4400 379855 7602466 P1 Schizomida Hubbardiidae 1 T138503 Molecular Potential SRE sp. ‘SCH003’ Paradraculoides TOBRC0023 379345 7599952 P2 Schizomida Hubbardiidae 1 T138571 Molecular Potential SRE sp. ‘SCH003’ Specimen likely Paradraculoides MEARC4273 382991 7602721 P1 Schizomida Hubbardiidae 1 T138502 Molecular from East sp. ‘SCH004/004a’ Deepdale Site Cryptops sp. Potential SRE / MEARC4383 381850 7602435 P2 Scolopendromorpha Cryptopidae 1 T138573 Molecular 'CHI002’ Singleton Symphyla sp. Potential SRE / MEARC4273 382991 7602721 P1 Symphyla Scolopendrellidae 1 T138524 Molecular 'SYM025' Singleton Nicoletiinae sp. Widespread MEARC4383 381850 7602435 P2 Thysanura Nicoletiidae 1 90775 Molecular 'TN010' Species Subnicoletiinae MEARC3814 379907 7605517 P1 Thysanura Nicoletiidae 1 90768 Molecular Potential SRE sp. 'TS005'

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev 0.docx Warramboo Troglofauna Assessment

Appendix 3

Desktop Review: Troglobitic Fauna Species Lists and Conservation Significance

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Number Site Collection Order Family Species ID method Species status Collected Year Araneae Oonopidae Oonopidae sp. 'AO014' 1 Molecular DCBRC_017 2010 Potential SRE Araneae Oonopidae Oonopidae sp. 'AO014' 1 Molecular DCBRC_040 2010 Potential SRE Araneae Oonopidae Prethopalpus sp. 'B21' 1 Morphological DCBRC_089 2010 Potential SRE Araneae Oonopidae Prethopalpus sp. 'B21' 2 Morphological MEARC4191 2010 Potential SRE Cephalostigmata Indeterminate Symphyla sp. 'SYM025' 1 Molecular TOBRC0038 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0003 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0009 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0010 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0030 2010 Potential SRE Cephalostigmata Scutigerellidae Hanseniella sp. 'B10' 1 Morphological COBRC0033 2010 Potential SRE Coleoptera Curculionidae Curculionidae sp. 'B10' 2 Morphological COBRC0007 2010 Potential SRE Diplura Japygidae Japygidae sp. 'B17' 2 Morphological COBRC0008 2010 Potential SRE Diplura Parajapygidae Parajapygidae sp. 'B14' 1 Morphological COBRC0003 2010 Potential SRE Diplura Projapygidae Projapygidae new genus sp. 1 Morphological MEA4318 2007 Potential SRE ‘nov.’ Diplura Projapygidae Projapygidae sp. 'B6' 1 Morphological DCBRC_001 2010 Potential SRE Diplura Projapygidae Projapygidae sp. 'B6' 3 Morphological TOBRC0043 2010 Potential SRE Isopoda Armadillidae Armadillidae sp.’ISA009a/9b’ 1 Molecular MEARC5044 2013 Widespread species Isopoda Armadillidae Armadillidae sp.’ISA009a/9b’ 1 Molecular TOBRC0003 2013 Widespread species Isopoda Armadillidae Armadillo sp. 'B18' 2 Morphological DCBRC_0038 2010 Potential SRE Isopoda Armadillidae Armadillo sp. 'B18' 2 Morphological DCBRC_089 2010 Potential SRE Isopoda Armadillidae Armadillo sp. 'B18' 1 Morphological TOBRC0044 2010 Potential SRE Isopoda Philosciidae Philosciidae sp. 'ISP051' 1 Molecular RC12COB0002 2013 Potential SRE Palpigradi Indeterminate Palpigradi sp. 'B6' 3 Morphological COBRC0022 2010 Potential SRE Polyxenida Lophoproctidae Lophoturus madecassus 1 Morphological COBRC0022 2013 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 1 Morphological MEA2999 2007 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 2 Morphological MEA4318 2007 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 9 Morphological MEARC4329 2013 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 2 Morphological MEARC5038 2013 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 36 Morphological MEARC5044 2013 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 11 Morphological TOBRC0009 2013 Widespread species Pseudoscorpiones Atemnidae Atemnidae sp. 'B2' 1 Morphological DCBRC_017 2010 Potential SRE Pseudoscorpiones Chthoniidae Tyrannaochthonius sp. 1 Morphological MEADC2380 2005 Potential SRE 'Warramboo'

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Number Site Collection Order Family Species ID method Species status Collected Year Pseudoscorpiones Chthoniidae Tyrannochthonius basme 2 Morphological COBRC0010 2010 Potential SRE Pseudoscorpiones Olpiidae Olpiidae sp. 'POL013' 1 Molecular COBRC0013 2010 Potential SRE Pseudoscorpiones Syarinidae Ideoblothrus linnaei 1 Morphological MEA4316 2007 Conservation significant, and Molecular Listed as Priority 1 in WA Pseudoscorpiones Syarinidae Ideoblothrus sp. 'Mesa A' 1 Morphological MEA4063 2007 Conservation significant, Listed as Priority 1 in WA Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEA3092 2006 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 2 Morphological MEADC2492 2007 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEADC3188 2007 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEARC2702 2005 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEARC2740 2014 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEARC3066 2005 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEARC3073 2005 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Draculoides anachoretus 1 Morphological MEARC4151 2005 Conservation significant, and Molecular Listed as Schedule 3 in WA. Schizomida Hubbardiidae Paradraculoides sp. ‘SCH003’ 4 Morphological MEADC2381 2005 Potential SRE and Molecular Scutigerida Cryptopidae Cryptopidae sp. 'CHI022' 1 Molecular COBRC0001 2010 Potential SRE Scutigerida Cryptopidae Cryptops sp. 'B14' 1 Morphological COBRC0019 2010 Potential SRE Zygentoma Indeterminate Thysanura sp. 'T001' 1 Molecular TOBRC0055 2013 Potential SRE Zygentoma Nicoletiidae Atelurodes sp. 'B2' 8 Morphological DCBRC_017 2010 Potential SRE Zygentoma Nicoletiidae Atelurodes sp. 'B2' 4 Morphological DCBRC_089 2010 Potential SRE

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Number Site Collection Order Family Species ID method Species status Collected Year Zygentoma Nicoletiidae Hemitrinemura sp. 'B4' 1 Morphological COBRC0007 2010 Potential SRE Zygentoma Nicoletiidae Hemitrinemura sp. 'B4' 2 Morphological COBRC0016 2010 Potential SRE Zygentoma Nicoletiidae Trinemura sp. 'B10' 10 Morphological MEARC4191 2010 Potential SRE Zygentoma Nicoletiidae Trinemura sp. 'B11' 1 Morphological COBRC0019 2010 Potential SRE Zygentoma Nicoletiidae Trinemura sp. ‘Mesa A1’ 2 Morphological MEARC3098 2005 Potential SRE Zygentoma Nicoletiidae Trinemura sp. ‘T1’ 2 Morphological TOBRC0011 2013 Potential SRE Cephalostigmata Indeterminate Symphyla sp. 'SYM025' 1 Molecular TOBRC0038 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0003 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0009 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0010 2010 Potential SRE Cephalostigmata Scolopendrillidae Symphylella sp. 'B9' 1 Morphological COBRC0030 2010 Potential SRE Cephalostigmata Scutigerellidae Hanseniella sp. 'B10' 1 Morphological COBRC0033 2010 Potential SRE Coleoptera Curculionidae Curculionidae sp. 'B10' 2 Morphological COBRC0007 2010 Potential SRE Diplura Japygidae Japygidae sp. 'B17' 2 Morphological COBRC0008 2010 Potential SRE Diplura Parajapygidae Parajapygidae sp. 'B14' 1 Morphological COBRC0003 2010 Potential SRE Diplura Projapygidae Projapygidae new genus sp. 1 Morphological MEA4318 2007 Potential SRE ‘nov.’ Diplura Projapygidae Projapygidae sp. 'B6' 1 Morphological DCBRC_001 2010 Potential SRE Diplura Projapygidae Projapygidae sp. 'B6' 3 Morphological TOBRC0043 2010 Potential SRE Isopoda Armadillidae Armadillidae sp.’ISA009a/9b’ 1 Molecular MEARC5044 2013 Widespread species Isopoda Armadillidae Armadillidae sp.’ISA009a/9b’ 1 Molecular TOBRC0003 2013 Widespread species Isopoda Armadillidae Armadillo sp. 'B18' 2 Morphological DCBRC_0038 2010 Potential SRE Isopoda Armadillidae Armadillo sp. 'B18' 2 Morphological DCBRC_089 2010 Potential SRE Isopoda Armadillidae Armadillo sp. 'B18' 1 Morphological TOBRC0044 2010 Potential SRE Isopoda Philosciidae Philosciidae sp. 'ISP051' 1 Molecular RC12COB0002 2013 Potential SRE Palpigradi Indeterminate Palpigradi sp. 'B6' 3 Morphological COBRC0022 2010 Potential SRE Polyxenida Lophoproctidae Lophoturus madecassus 1 Morphological COBRC0022 2013 Widespread species Polyxenida Lophoproctidae Lophoturus madecassus 1 Morphological MEA2999 2007 Widespread species

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Appendix 4

Helix Molecular Solutions: Genetic Analysis Reports

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx

Molecular systematics of subfauna from the Robe Valley

6 January, 2015

Jason Alexander Biota Environmental Sciences Level 1, 228 Carr Place Leederville, WA 6007

Via email

Re. Report on the molecular systematics of subfauna from the Robe River Valley

Dear Jason,

Following is a summary of the results of the subfauna study we have completed on nine taxonomic groups from the Robe River Valley. Fifty-five distinct genetic lineages were detected among the seven groups for which sequences were obtained. The 55 lineages likely represent between 51 and 54 species, only five of which have been detected previously in the Pilbara. A further species matched a Genbank specimen of Coleoptera which does not appear to have been recorded in Australia, although the family to which it belongs is well-represented.

Thanks once again for collaborating on this project with Helix. We hope we can continue to provide you with useful information, and feel free to contact us if you have any questions or would like to discuss the results in detail.

Sincerely,

Dr. Terrie Finston, Yvette Hitchen and Dr. Oliver Berry Helix Molecular Solutions

1 Molecular systematics of subfauna from the Robe Valley

Background and Objective Two hundred and thirty specimens of subfauna (troglofauna and stygofauna) belonging to nine taxonomic groups (Amphipoda, Chilopoda, Coleoptera, Diplopoda, Diplura, Isopoda, Pseudoscorpiones, Schizomida, Thysanura) were collected from Robe River Valley and sequenced for variation at the mitochondrial COXI gene. The molecular data were assessed in order to determine the number of species present in each group and compare the results to those obtained during previous surveys that have been undertaken for the these groups elsewhere in the Pilbara.

Executive summary  Two hundred and thirty specimens from the Robe River Valley belonging to nine taxonomic groups were sequenced for variation at the COI gene.  Fifty-five lineages were detected among the seven groups for which sequences were obtained.  The 55 lineages likely represent between 51 and 54 species.  Five of the species have been detected previously in the Pilbara, whereas the remainder appear to be new, based on the material available for comparison.

Methods Two hundred and thirty-eight specimens of subfauna collected from the Robe River Valley area were sequenced for variation at the mitochondrial cytochrome oxidase subunit I gene (COXI) using multiple pairs of primers (LCOI/HCO2, LCOI-long/HCO2-long, NemF1/NemR1 and LCOI/CIN2341.

Sequences were edited using GENEIOUS software (Drummond et al. 2011). Alignment was performed with CLUSTAL W (Thompson et al. 1994) using default parameters. Genetic distances between unique genetic sequences (haplotypes) were measured using uncorrected p- distances (total percentage of nucleotide differences between sequences).

MODELTEST software (Posada and Crandall, 1998) was used to determine the model of sequence evolution that best fitted the data for each taxonomic group. Bayesian analysis was used to construct the phylogenetic trees, incorporating the model as identified in MODELTEST for each taxonomic group. The phylogeny, branch lengths and posterior probabilities were obtained by running two trees simultaneously, each running four simultaneous MCMC chains. The number of cycles needed was determined by the standard deviation of the split frequencies of the two trees. The analysis was paused after every 1 x 106 generations and when the standard deviation fell below 0.01, the analysis was stopped. A majority rule consensus tree was constructed after discarding the “burn-in” trees in both analyses. The burn-in value was determined by plotting the posterior probabilities obtained after every generation and identifying the point at which the values reach stationarity (= the asymptote). Trees produced prior to stationarity were discarded.

For the purposes of this report, lineages were defined as haplotypes or groups of haplotypes differing from other such groups by >3% sequence divergence. This cut-off was selected based on bar-coding data, which indicates that intra-specific variation rarely exceeds 3% (Hebert et al., 2003b).

Results Schizomids Preliminary analysis - Reference sequences and outgroups Ninety-one schizomids were sequenced from the Robe River Valley (Table 1). In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted with the 91 Robe River Valley sequences to identify the number of genetic lineages present. A representative from each lineage was analysed with 373 reference sequences of schizomids from previous surveys in the Pilbara to compile a manageable and relevant reference data set. The reference sequences were selected based on the criteria that they showed ≤ 15% sequence divergence from at least one of the Robe River Valley lineages.

2 Molecular systematics of subfauna from the Robe Valley

Ten distinct genetic lineages were identified (Figure 1) and a representative from each was analysed in a model-based phylogenetic analysis with 26 reference specimens. Two representatives were included from lineage 4 (lineage 4 and 4a) to more fully represent the genetic variation within the lineage. A specimen of schizomid from the family Hubbardiidae, Brignolozomus woodwardi (GenBank accession # EU272675) was used as an outgroup.

Phylogenetic analysis The phylogenetic analysis, which included the ten Robe River Valley lineages, in addition to 26 reference specimens of schizomids, placed the Robe River Valley specimens in ten distinct lineages, two of which (lineages 8 and 1) also contained reference specimens from a previous survey at the Robe River Valley (Draculoides mesozeirus and P. bythius). A third Robe Valley lineage (lineage 3) formed a close relationship with Paradraculoides sp. whereas the remaining seven lineages were genetically distinct from the reference specimens as evidenced by long branch lengths (Figure 2). Lineages 1 -4 formed a well-supported clade with five reference specimens of Paradraculoides and lineages 7 – 9 formed a well-supported clade with the reference specimen of Draculoides mesozeirus (Figure 2). Lineage 10 formed a well-supported clade with specimens identified as Draculoides from the Pilbara, although no Genbank voucher specimens were contained in the clade (Figure 2). Lineage 5 was highly distinct, and was placed outside all of the clades containing Draculoides and Paradraculoides specimens (Figure 2).

Differentiation within and between lineages Lineage 4 and 4a differed from one another by an average of 3.1% sequence divergence (Table 2). Excluding the divergence within lineage 4, the ten lineages of schizomids detected at the Robe River Valley differed from one another by between 5.8 and 18.0% mean sequence divergence (Table 2). Individuals within each of the ten lineages differed from one another by between 0.1 and 1.3% mean sequence divergence (Table 3).

Two of the ten lineages (8 and 1) differed from the closest reference lineage (D. mesozeirus and P. bythius, respectively), collected during previous surveys at the Robe River Valley by <1% sequence divergence (Table 4). Lineage 3 differed from the closest reference lineage (Paradraculoides sp.) by 3.9% (Table 4). The remaining lineages differed from the nearest reference specimens by >7% (Table 4).

Distribution of lineages Two lineages (1 and 4/4a) were detected in multiple deposits (Table 5). Lineage 1 was detected at Mesa B and C, and lineage 4/4a was detected at Mesas L, M and N, and Highway and Tod bore (Table 5). Two deposits (Mesa B and Highway and Tod bore) contained multiple lineages. Mesa B contained lineages 1 and 5, and Highway and Tod bore contained lineages 3 and 4 (Table 5).

Amphipoda Preliminary analysis - Reference sequences and outgroups Forty-seven specimens of amphipods from seven sites were sequenced from Robe Valley (Table 6). In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted on the 25 Robe Valley amphipods for which sequences were obtained, and haplotypes were selected from each genetic lineage to include in a model-based phylogenetic analysis. Two sequences of the cladoceran genus Daphnia were used as outgroups for both analyses as follows: Daphnia pulex (Genbank accession number J150976) and Daphnia pulicaria (Genbank accession number JN233925).

The neighbour-joining analysis placed the 25 specimens of amphipods for which sequences were obtained into eight lineages (Figure 3). Comparisons to the GenBank sequence database indicated that six of the eight lineages (1 – 6) were most similar to sequences of Melitidae, whereas two lineages (7, 8) were most similar to sequences of Niphargidae. The Melitidae lineages contained between one and six specimens, and the putative Niphargidae lineages contained between one and five specimens (Figure 3).

For the Melitidae, 20 reference sequences were included, two GenBank vouchers of Melitidae, Norcapensis mandibularis (Genbank accession #JQ608487) and Nedsia sp. (Genbank

3 Molecular systematics of subfauna from the Robe Valley

accession # EU304458) and 18 specimens of Melitidae detected in previous surveys of the Robe River, Cape Range, Barrow Island and Ashburton River drainage basins.

For the Niphargidae, two reference sequences were included in the phylogenetic analysis, , Niphargus fontanus (Genbank accession #KC315635) and Niphargus glenniei (Genbank accession # KC315646) as well as 19 specimens from previous surveys of the Central Pilbara, Barrow Island, Lower Shaw River, Fortescue River, and Marillana Creek.

Phylogenetic analysis The phylogenetic analysis, revealed the presence of two distinct, well-supported clades, one containing lineages 1-6 and reference specimens of Melitidae and one containing lineages 7 and 8 and reference specimens of Niphargidae (Figure 4). Two of the two Robe Valley Melitidae lineages contained closely related reference specimens. Lineages 1 (IV126) and 2 (IV92) formed well-supported clades with reference specimens of Nedsia from a previous survey in the Robe River Valley, rr3a and nr5mi well, respectively (Figure 4). Further, lineage 5 formed a well-supported clade with specimens of Nedsia from Barrow Island and the Cape Range peninsula, and lineages 7 and 8 formed a well-supported clade with each other and the two Niphargus reference sequences (Figure 4).

Differentiation within and between lineages The six lineages of Melitidae detected at Robe Valley differed from one another by between 6.7 and 15.6% mean sequence divergence and the two lineages of Niphargidae differed by 15.1% sequence divergence (Table 7). Differentiation within the six melitid lineages averaged between 0.0 and 1.8% sequence divergence and differentiation within Niphargidae lineage 8 averaged 0.5% sequence divergence (Table 8).

Robe Valley Melitidae lineage 1 differed from the closest reference lineage (rr3a-2b) by 2.7% and Melitidae lineage 2 differed from the closest reference lineage (nr5miwell-d) by 0.2% (Table 9). The remaining lineages of Melitidae differed from the nearest reference lineages by between 6.8 and 10.3% sequence divergence (Table 9). The two lineages of Niphargidae from the Robe Valley differed from the reference lineages of Niphargus by between 19.0 and 21.3% sequence divergence (Table 9).

Distribution of lineages Three of the eight lineages were detected at multiple sites (Table 10), Similarly, three of the six sites contained multiple lineages- lineages 1, 2 and 8 were detected at Budgie, lineages 4, 7 and 8 were detected at Daves and lineages 3 and 5 were detected at MB13WARR (Table 10).

Chilopoda Five specimens of Chilopoda from three deposits were sequenced (Table 11). Sequences were obtained from four specimens and these were analysed with six Genbank vouchers from four families of Chilopoda, Cryptopidae, Henicopidae, Geophilidae and Mecistocephalidae (in the superfamily Geophilomorpha) as follows: Scolopocryptops sexspinosus (Genbank accession # AY288745), Paralamyctes monteithi (Genbank accession # AF334321), Gnathoribautia bonensis (Genbank accession # KF569297), Geophilus flavus (Genbank accession # JN306685), Stenotaenia linearis (Genbank accession # KR736251), and Mecistocephalus multidentatus (Genbank accession # AB610774) as well as 17 reference specimens of Chilopoda from nine sites in the Pilbara. Two specimens of Onychophora, Metaperipatus inae (Genbank accession # HQ453464) and Opisthopatus cinctipes (Genbank accession #NC014273) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the four specimens from the Robe Valley in addition to 23 reference specimens, placed the Robe Valley specimens in two genetically distinct clades (Figure 5). Three of the specimens (IV139- IV141) formed a well-supported lineage, which was placed in a well-supported clade with Genbank specimens of Geophilomorpha (Geophilidae and Mecistocephalidae; Figure 5). The fourth specimen (IV143) was placed in a well-supported

4 Molecular systematics of subfauna from the Robe Valley

clade containing reference specimens from the Pilbara assigned to the genus Cryptops (Figure 5).

Differentiation within and between lineages Robe Valley Chilopoda lineage 1 (specimens IV139 – IV141) differed from one another by between 0.0 and 0.3% sequence divergence and from lineage 2 (IV143) by 22.8 – 22.9% sequence divergence (Table 12). The Robe Valley specimens differed from the reference specimens by between 18.0 and 28.1% sequence divergence (Table 12).

Distribution of lineages The Robe Valley Chilopoda lineage 1 was detected at deposit 2403A and lineage 2 was detected at Highway/Tod bore (Table 13). Coleoptera Eleven specimens of Coleoptera from five drillholes at four sites were sequenced from the Robe Valley (Table 14). The nine specimens for which sequences were obtained were analysed with six Genbank voucher sequences of Coleoptera, selected on their similarity to the Robe Valley specimens: Stricticollis tobias (Anthicidae; Genbank accession # KJ962132), Necrobia violacea (Cleridae; Genbank accession #KJ961916), Pterostichus mutus (Carabidae; Genbank accession #EU710801), Harpalus discrepans (Carabidae; HM180603), Trigonopterus sp (Curculionidae; Genbank accession #HE615891), Echinodera ibleiensis (Curculionidae, Genbank accession #GU213687) as well as 14 reference specimens of Coleoptera from six sites in the Pilbara. Two specimens of Hemiptera (Nisia carolinensis Genbank accession #LC015451) and Lycorma delicatula (Genbank accession #EU909203) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the nine specimens from the Robe Valley for which sequences were obtained in addition to 20 reference specimens, placed the Robe Valley specimens in six genetically distinct lineages (Figure 9). The lineages contained between one and four specimens (Figure 6). One of the Robe Valley lineages contained a closely related reference specimen. Lineage 3 (IV153) contained the reference specimen of Stricticollis tobias from the family Anthicidae (Figure 6). The remaining lineages did not show close relationships with other reference specimens, however, most of them were placed in well-supported clades containing reference specimens, allowing some tentative taxonomic assignments. Lineage 1 (IV151) formed a well-supported clade with specimens of Carabidae from Genbank and the Pilbara, and lineages 4 and 5 (IV150 and IV152, respectively)formed a well-supported clade with each other and the Genbank and reference sequences of Curculionidae (Figure 6). Lineage 6 formed a well-supported clade containing Genbank specimens of Curculionidae as well as Pilbara specimens of Curculionidae (Figure 9). The remaining lineage was less well resolved. Lineage 2 formed a well-supported clade with Genbank reference specimens of both Anthicidae and Cleridae (Figure 6).

Differentiation within and between lineages The four specimens within lineage 2 were genetically identical (Table 15). The six Robe Valley lineages differed from one another by between 3.2 and 27.1% sequence divergence (Table 15). The Robe Valley lineages differed from the reference specimens by between 0.9 and 31.0% sequence divergence (Table 15).

Distribution of lineages The Robe Valley Coleoptera lineages were each detected in singe deposits (Table 16). Similarly, each deposit contained a single lineage, except Highway/Tod which contained three lineages: 1, 3 and 5 (Table 16).

Diplopoda

5 Molecular systematics of subfauna from the Robe Valley

Reference specimens and outgroups Eleven specimens of Diplopoda assigned to the Polydesmida on the basis of morphology, were sequenced (Table 17). Six specimens yielded a DNA sequence, however, the DNA appeared to be contaminated with DNA from flies (muscomorpha).

Diplura Reference specimens and outgroups Seven specimens of Diplura from six drillholes at three deposits were sequenced (Table 18). The five specimens for which sequences were obtained were analysed with 20 specimens of Diplura from nine sites in the Pilbara belonging to four families: Anajapygidae, Japygidae, Parajapygidae and Projapygidae, as well as four Genbank voucher specimens, Diplura sp. (Genbank accession #HQ943342), Campodea tillyardi (Genbank accession # AF370844) and Lepiodocampa weberi (Genbank accession #HQ882832) from the family Campodeidae and Japyx solifugus (Genbank accession #AY771989) from the family Japygidae. The planthopper Lycorma delicatula (Genbank accession # FJ456942) and a specimen of Fulgoridae (Hemiptera sp.; Genbank accession #GU671563) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the five specimens of Diplura from Robe Valley in addition to 24 reference specimens, placed the Robe Valley specimens in four distinct lineages (Figure 7). The Robe Valley specimens showed no close relationship to any of the reference specimens, however, one (lineage 4) was placed in a well-supported clade containing the Pilbara specimens of Parajapygidae and three (lineages 1, 2 and 3) were placed in a well- supported clade containing the Genbank reference specimens of Campodeidae (Figure 7).

Differentiation within and between lineages The two specimens from the Robe Valley in lineage 3 differed from one another by 0.6% sequence divergence (Table 19). The Robe Valley lineages of putative Campodeidae differed from one another by between 4.7 and 15.3% sequence divergence and from the nearest reference specimen (Campodea tillyardi) by 21.4 to 22.2% sequence divergence (Table 19). The Robe Valley specimen of Parajapygidae differed from the nearest reference specimen (BX29 from Murrays Hill) by 17.5% sequence divergence (Table 19). Distribution of lineages The Robe Valley Diplura lineages were each detected in singe deposits (Table 20). Mesa C contained a single lineage (lineage 4), however Mesa B contained three lineages: 1, 2 and 3 (Table 20). Two lineages, 1 and 2, were found in the same drillhole at Mesa B (Table 20).

Isopoda Preliminary analysis, reference specimens and outgroups Twenty-seven specimens of Isopoda were sequenced from 17 drillholes at six deposits at Robe Valley (Table 21). Fifteen specimens yielded a DNA sequence. In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted on the 15 Robe Valley isopods for which sequences were obtained, and haplotypes were selected from each genetic lineage to include in a model-based phylogenetic analysis. Ten distinct genetic lineages were detected in the NJ analysis (Figure 8), and the ten representatives were analysed in a model-based phylogenetic analysis with 35 reference specimens of both aquatic and terrestrial Isopoda from Genbank and 14 sites in the Pilbara. While the specimens were collected from troglofauna traps, a search of similar sequences on Genbank and in the Helix database indicated the specimens were most closely matched with Haloniscus, Burmoniscus and Armadillidium, the former of which is an aquatic genus. It should be noted however, that Haloniscus is considered to be a terrestrial isopod living in aquatic habitats (Bayley and Ellis, 1969). Hence we included Genbank voucher specimens of both aquatic and terrestrial isopods: Pygolabis sp. 1 (Genbank accession # EU107646) and Pygolabis

6 Molecular systematics of subfauna from the Robe Valley

sp. 4 (Genbank accession #EU107664), Haloniscus sp. 10 (Genbank accession #EU364592), Halonsicus sp. 12 (Genbank accession #EU364601), Halonsicus sp. 21 (Genbank accession #EU364622) Laevophiloscia yalgoorensis (Genbank accession #EU364629), Armadillidium nasatum (Genbank accession #FN824099) and Burmoniscus sp. (Genbank accession #AB626254). Two specimens of the shrimp Stygiocaris, S. stylifera (Genbank accession #EU123818) and S. lancifera (Genbank accession #EU123826) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the ten representatives from Robe Valley in addition to 35 reference specimens, placed the Robe Valley specimens in ten distinct lineages (Figure 9). One of the lineages (lineage 9a; IV181) also contained a reference specimen from Mesa A (Figure 9). Lineage 9a in turn, formed a well-supported clade with lineage 8 and 9b (Figure 9). The remaining lineages did not show close relationships with any of the reference specimens, however lineages 1 and 2 formed a well-supported clade with one another and with two lineages from Buckland Hills and Red Hill (Figure 9). Lineages 3 and 4 formed a poorly supported clade with the clade containing lineages 1 and 2 (Figure 9). Lineages 5, 6, and 7 formed a well- supported clade with specimens of Troglarmadillo from the Pilbara (Figure 9).

Differentiation within and between lineages The Robe Valley lineages of Isopoda differed from one another by a mean of 3.7 to 25.9% sequence divergence (Table 22). Differentiation within lineages ranged from a mean of 0.1 to 2.5% sequence divergence (Table 23). The Robe valley lineages differed from the reference specimens by between 1.2 and 30.1% sequence divergence (Table 24). In particular, lineages 9a and 9b differed from IE45 (a Mesa A reference specimen) by 1.2 to 3.8% sequence divergence (Table 24). The remaining Robe Valley lineages differed from the reference lineages by >12% (Table 24). Distribution of lineages The Robe Valley Isopoda lineages were each detected in singe deposits (Table 24). Mesa B contained three lineages: 4, 5 and 8, and Highway/Tod bore contained four lineages: 6, 7, 9a and 9b (Table 25). One drillhole, TOBRC0020 at Highway/Tod bore contained two lineages, 6 and 7 (Table 25).

Pseudoscorpions Reference sequences and outgroups Twenty-six pseudoscorpions were sequenced from 16 drillholes at seven deposits at Robe Valley (Table 26). In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted on the 18 Robe Valley pseudoscorpions for which sequences were obtained, and haplotypes were selected from each genetic lineage to include in a model-based phylogenetic analysis. Fifteen distinct genetic lineages were detected in the NJ analysis (Figure 10), and the 15 representatives were analysed in a model-based phylogenetic analysis with selected reference specimens. A search of similar sequences on Genbank and in the Helix database indicated that four of the lineages (9 – 12) most closely matched the family Chthoniidae and 11 of the lineages most closely matched Helix database specimens of Hyiidae and a Genbank specimen of Atemnidae. Based on those results, GenBank voucher sequences were included as reference sequences of Hyiidae (one specimen) and Atemnidae (one specimen) as follows: Indohya sp JM 2008 (Genbank accession # EU559564) and Paratemnoides sumatranus (GenBank accession # JN018204). In addition, as local references, seven specimens of Hyiidae from seven sites in the Pilbara and 22 specimens of Chthoniidae from 11 sites were included in the analysis. Sequences of the scorpion Pandinus imperator (Genbank accession # AY1565821) and harvestman spider Siro rubens (Genbank accession # DQ5131111) were used as outgroups.

Phylogenetic analyses Two phylogenetic analyses were conducted, one for the Chthoniidae and one for the Hyiidae/Atemnidae. The phylogenetic analysis of the Chthoniidae, which included the four Chthoniidae lineages from Robe Valley, in addition to 25 reference specimens, placed the

7 Molecular systematics of subfauna from the Robe Valley

Robe Valley specimens in four distinct genetic lineages (Figure 11). The Robe Valley Chthoniidae lineages did not show close relationships to any of the reference specimens, however lineages 9 and 10 formed a well-supported clade with one another and with six specimens of Chthoniidae from the Red Hill and Ophthalmia (Figure 11). Similarly, lineages 11 and 12 formed a well-supported clade with one another and with a specimen of Chthoniidae from Red Hill (Figure 11).

The phylogenetic analysis of the Hyiidae and Atemnidae, which included the 11 lineages from Robe Valley, in addition to nine reference specimens, placed the Robe Valley specimens in 11 distinct genetic lineages (Figure 12). Lineages 1 – 8 were placed in a clade containing the Pilbara and Genbank reference specimens of Hyiidae (Figure 12). The Robe Valley Hyiidae lineages did not show close relationships to any of the reference specimens, however lineages 3 and 4 formed a well-supported clade with one another and with two specimens of Hyiidae from the Central Pilbara and Hardey River (Figure 12). Similarly, lineages 6, 7 and 8 formed a well-supported clade with one another and with a specimen of Hyiidae from Buckland Hills (Figure 12). The remaining Hyiidae lineages were poorly resolved (Figure 12). Lineages 13 – 15 formed a well-supported clade with one another and with the Genbank specimen of Atemnidae (Figure 12).

Differentiation within and between lineages The fifteen lineages of pseudoscorpions from the Robe Valley differed from one another by between a mean of 3.8 and 29.5% sequence divergence (Table 27). Variation within lineages ranged from between a mean of 0.1 and 1.2% sequence divergence (Table 28). The four lineages of Chthoniidae from the Robe Valley differed from one another by between a mean of 10.5 and 22.6% sequence divergence (Table 27), and from the reference specimens of Chthoniidae by between 14.4 and 29.6% sequence divergence (Table 29). The eight lineages of Hyiidae from the Robe Valley differed from one another by between a mean of 3.8 and 26.5% sequence divergence (Table 27), and from the reference specimens of Hyiidae by between 11.3 and 34.9% sequence divergence (Table 30). The three lineages of Atemnidae differed from one another by between a mean of 7.6 and 14.6% sequence divergence (Table 27) and from the reference specimens of Atemnidae by between 18.0 and 19.8% sequence divergence (Table 30).

Distribution of lineages The Robe Valley pseudoscorpion lineages were each detected in singe deposits (Table 31). Only two of the 15 lineages were detected in more than one bore – Chthoniidae lineage 11 and Atemnidae lineage 15 were both detected in two bores (Table 31). Deposits Mesa B, 2402D and 2403A each contained three lineages and Highway/Tod bore and 2402E each contained two lineages, whereas two deposits, Mesa L and 2401A contained single lineages (Table 31).

Thysanura Reference specimens and outgroups Four specimens of Thysanura were sequenced from three bores at two deposits (Table 32). None of the specimens yielded a DNA sequence.

8 Molecular systematics of subfauna from the Robe Valley

data). Similarly, the four described species of Draculoides differ from one another by between 4.5 to 13.7% sequence divergence (uncorrected p-distances calculated by us from Harvey et al., 2008).

Ten genetically distinct lineages of schizomids were detected at the Robe River Valley. The ten lineages differed from one another by >approximately 6% sequence divergence, thus each is likely to represent a distinct species. Variation within lineage 4 (between lineages 4 and 4a) is slightly higher than would be expected within a single species, and this is likely due to geographic separation between Mesa L, where lineage 4a was detected, and Highway and Tod bore and Mesas M and N, where lineage 4 was detected. Thus lineages 4 and 4a may represent incipient species, or a single species showing differentiation due to limited sampling or the poor dispersal ability of the group. We suggest the latter conclusion of a single species is more likely, however, sampling additional specimens from intermediate areas may help resolve this.

Two of the lineages, 1 and 8, corresponded to previously described species, P. bythius and D. mesozeirus, respectively. A third lineage (3) may correspond to a third described species, Paradraculoides sp. OFB 2008, although the genetic distance between the specimens from the present study and the reference specimen was somewhat higher than is generally observed between individuals of the same species. Thus lineage 3 and Paradraculoides sp. may represent incipient species, or a single species showing differentiation due to limited sampling or the poor dispersal ability of the group. Morphological evidence or sampling additional specimens from intermediate areas may help resolve this relationship. The remaining seven lineages appear to be new, based on the material available for comparison. Lineages 1 – 4 are likely to belong to the genus Paradraculoides and lineages 7 – 9 are likely to belong to the genus Draculoides. Lineage 10 may also belong to the genus Draculoides, although more voucher sequences are needed for this genus.

Amphipoda Eight genetically distinct lineages of amphipods were detected at the Robe River Valley. The eight lineages differed from one another by > 6% sequence divergence, thus each is likely to represent a distinct species. Six of the species are Melitidae, and two show the greatest similarity to sequences of Niphargidae. Lineages 1-5 were placed in the large clade containing reference specimens of Nedsia and thus are all likely to belong to that genus. Lineage 6 was placed outside the Nedsia clade and may represent a different genus of Melitidae. Two of the Nedsia species have been detected previously in the Robe Valley (lineages 1 and 2), based on their high genetic similarity to reference sequences (<3% sequence divergence). In contrast, the remaining four Melitidae lineages and the two Niphargidae lineages differed from the nearest reference lineages by >7% sequence divergence and thus appear to be new, based on the material available for comparison.

Chilopoda Two genetically distinct lineages of Chilopoda were detected at the Robe River Valley. The two lineages differed from one another by > 20% sequence divergence, thus each is likely to represent a distinct species. Neither showed close genetic relationships to reference specimens, differing from the references by >18.0% sequence divergence, thus both species are likely to be new, based on the material available for comparison. Lineage 1 was placed with the Geophilomorpha, and lineage 2 was placed with specimens of the genus Cryptops.

Coleoptera Six genetically distinct lineages of Coleoptera were detected at Robe Valley. Two of the lineages, 4 and 5, differed from one another by 3.2% sequence divergence. As they were detected at different deposits, it is likely that they represent a single species, which shows genetic differentiation due to the geographic distance between deposits. The remaining lineages differ from one another by ≥ 19% sequence divergence, thus each is likely to represent a distinct species. With one exception, the lineages detected at Robe Valley differ from the reference lineages by >10% sequence divergence, thus each is likely to represent a new species, which has so far not been detected, based on the material available for comparison. Only lineage 3 showed a close relationship to a reference specimen. Lineage 3 showed <1% sequence divergence from the Genbank specimen of Stricticollis tobias in the family Anthicidae

9 Molecular systematics of subfauna from the Robe Valley

and the subfamily Anthicinae. While we could find no record of this species in Australia, the subfamily is well-represented, with 18 genera recorded from Australia (Australian Faunal Directory).

Diplopda None of the Diplopoda specimens yielded a useable DNA sequence.

Diplura Four distinct genetic lineages of Diplura were detected at Robe Valley. The four lineages differed from one another by a mean of >5%, thus each is likely to represent a distinct species. While the genetic distance between lineages 2 and 3 is moderately low (mean=5%), both lineages were detected at Mesa B, although in different drillholes, indicating there are barriers to gene flow between the two lineages over short geographic distances, supporting the notion that they are separate species. Sampling of intermediate sites may help resolve the relationship between lineages 2 and 3. All four lineages differ from the reference specimens by >20% and thus appear to be new, based on the material available for comparison. Three of the lineages (1 – 3) may belong to the Dipluran family Campodeidae, which is represented in Australia by four genera, and one lineage (4) likely belongs to the Parajapygidae family.

Isopoda Ten distinct genetic lineages of Isopoda were detected at Robe Valley. Two of the lineages, 9a and 9b differed from one another by a mean of 3.7%, which, while low, is higher than is typically observed between members of the same species. Both lineages were detected at the Highway/Tod bore deposit, although in different drillholes, indicating there are barriers to gene flow between the two lineages over short geographic distances, re-enforcing the notion of separate species. Sampling of intermediate sites may help resolve the relationship between lineages 9a and 9b. One of the lineages, 9b, was detected previously at Mesa A. The remaining lineages differ from the reference specimens by >12% and thus appear to be new, based on the material available for comparison. Three of the lineages (5 - 7) may belong to the genus Troglarmadillo, as they were placed in a large clade containing reference specimens of that genus, whereas the remaining lineages could not be given taxonomic assignments, owing to their distant relationships to reference taxa.

Pseudoscorpiones

Fifteen distinct genetic lineages of pseudoscorpions were detected at Robe Valley, four Chthoniidae, eight Hyiidae and three putative Atemnidae. The four lineages of Chthoniidae differed from one another by >10% sequence divergence, thus each is likely to represent a distinct species. Further, the four Chthoniidae lineages from Robe Valley differed from the reference specimens by >14%, indicating that the four are likely to be new species that have so far not been detected in the Pilbara, based on the material available for comparison.

The three putative lineages of Atemnidae differed from one another by >7% sequence divergence, thus each is likely to represent a distinct species. There are currently no DNA sequences of Atemnidae from other sites in the Pilbara available for comparison, thus we assign it tentatively to this family, and no conclusions can be reached regarding the broader distributions of these species. However, a new species of Anatemnus was recently described from the Pilbara, and four other species in the family are known from Australia (Alexander et al., 2014).

With two exceptions, the eight lineages of Hyiidae differed from one another by >13% sequence divergence. Two of the lineages, 3 and 4, differed from one another by a mean of 3.8%, which, while low, is higher than is typically observed between members of the same species. The two lineages were detected at different deposits, 2402D and 2402E, thus the observed genetic differentiation may reflect the poor dispersal abilities of this group. Sampling of intermediate sites may help resolve the relationship between lineages 3 and 4. In contrast, lineages 1 and 2 differed by approximately 5% sequence divergence, but both were detected at Mesa B, indicating that there are barriers to gene flow, over a relatively short geographic distance, and supporting the notion that the two should be considered separate species. However, sampling of intermediate sites may help resolve the relationship between the two lineages. The eight

10 Molecular systematics of subfauna from the Robe Valley

lineages differ from the reference specimens by >13% and thus appear to be new, based on the material available for comparison.

Summary Fifty-five lineages from seven taxonomic groups were detected at Robe Valley, as shown below. The 55 lineages likely represent 51 - 54 species. Five of the species have been detected previously in the Pilbara, and one matched a Genbank sequence of a species apparently not recorded from Australia, although the family to which it belongs is well-represented. The remaining species at Robe Valley are likely to be new, based on the material available for comparison. The failure rate was somewhat higher than usual with this project. In addition to our inability to produce amplification products from some specimens (e.g. Thysanura), of those that did amplify, some sequences were contaminated with foreign DNA - flies, even human. This happens when there isn't enough target DNA to out-amplify contaminants.

Schizomida Ten lineages were detected at Robe Valley, likely corresponding to ten species. Two of the species have been detected previously in the Pilbara (P. bythius and D. mesozeirus), and a third shows moderate genetic similarity to Paradraculoides sp. OFB 2008 and requires further investigation, while the remaining seven appear to be new, based on the material available for comparison.

Amphipoda Eight lineages were detected at Robe Valley, likely corresponding to eight species. Two of the species have been detected previously in the Pilbara (undescribed Melitidae, possibly Nedsia), while the remaining six appear to be new, based on the material available for comparison.

Chilopoda Two lineages were detected at Robe Valley, likely corresponding to two species. Neither of the species has been detected previously in the Pilbara, based on the material available for comparison.

Coleoptera Six lineages were detected at Robe Valley, likely corresponding to five species. One of the species matched a Genbank sequence of a species apparently not recorded from Australia (Stricticollis tobias from the family Anthicidae), although the family is well-represented in Australia. The remaining four species are likely to be new, based on the material available for comparison.

Diplopoda No clean sequences were obtained from the specimens of Polydesmida.

Diplura Four lineages were detected at Robe Valley, likely corresponding to four species. None of the species have been detected previously in the Pilbara, based on the material available for comparison.

Isopoda Ten lineages were detected at Robe Valley, likely corresponding to nine or ten species. One of the species has been detected previously at Mesa A, whereas the remainder appear to be new, based on the material available for comparison.

Pseudoscorpiones Fifteen lineages were detected at Robe Valley, likely corresponding to 13 - 15 species. None of the species have been detected previously in the Pilbara, based on the material available for comparison.

Thysanura No sequences were obtained from the specimens of Thysanura.

11 Molecular systematics of subfauna from the Robe Valley

References

Alexander, J. B. , Burger M. A.A., and Harvey, M.S. (2014). A new species of troglobitic Anatemnus (Pseudoscorpiones: Atemnidae) from the Pilbara bioregion of Australia. Records of the Western Australian Museum 29: 141 – 148.

Australian Faunal Directory. http://www.environment.gov.au/biodiversity/abrs/online- resources/. Accessed 27 Dec., 2015. Bayly, I. A.E, Ellis p. (1969). Haloniscus searlei chilton: An aquatic “terrestrial” isopod with remarkable powers of osmotic regulation. Comparative Biochemistry and Physiology 31: 523-528

Harvey, M. S., Berry, O. Edward, K. L., Humphreys, G. (2008) Molecular and morphological systematics of hypogean schizomids (Schizomida: Hubbardiidae) in semiarid Australia. Invertebrate Systematics 22: 167-194.

Hebert, P.D.N., Cywinska, A., Ball, S.L., deWaard J.R. (2003a). Biological identifications through DNA barcodes. Proceedings of the Royal Society of London B 270: 313-321. Hebert, P.D.N., Ratnasingham, S., deWaard J.R. (2003b). Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London B (supplement) 270: S96-S99. Helix Molecular Solutions (2015). Report on the molecular systematics of Schizomida from The Robe River Valley. Prepared for Biota Environmental Sciences, 5 February. Posada, D., Crandall, K.A. (1998). MODELTEST: testing the model of DNA substitution. Bioinformatics 14: 817-818.

Thompson, J., Higgins, D., and Gibson, T. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22: 4673–4680. doi:10.1093/nar/ 22.22.4673

12 Molecular systematics of subfauna from the Robe Valley

Table 1. Specimens of Schizomida used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking Number Cap # Helix ID Lineage P1 2402eUnk03-20150809-T1-01 343 IV01 Lin 8= D. mesozeirus P1 2402EUNK05T3-01 672 IV02 Lin 8= D. mesozeirus P1 2402eUnk08-20150809-T1-01 299 IV03 Lin 8= D. mesozeirus P1 2402eUNK09-T3-01 651 IV04 Lin 8= D. mesozeirus P1 2402eUnk11-20150809-T2-01 293 IV05 Lin 8= D. mesozeirus P1 2402EUNK13T2-01 692 IV06 Lin 8= D. mesozeirus STYGO P1 Budgie20150604-06 CAP 11 IV07 Lin 2 P1 DD11MEB001T3-02 683 IV08 Lin 1= P. bythius P1 DD14MEL0001-t2-01 759 IV09 Lin 4a P1 DD14MEM0001T2-03 525 IV10 Lin 4 P1 DD14MEN0001-20150808-T2-02 580 IV11 Lin 4 P1 DD14MRR0004-t2-01 782 IV12 Lin 7 P1 DD14MRR0005-T1-01 604 IV13 Lin 7 P1 DD14MRR0005-T1-01 IV14 Lin 7 P1 DD14MRR0005-T2-03 775 IV15 Lin 7 P1 DD14MRR007-20150808-T2-01 439 IV16 Lin 6 P1 DD14MRR007-20150808-T1-02 446 IV17 Lin 6 P1 DD14MRR0008-20150808-T3-01 494 IV18 Lin 6 P1 DD14MRR009-20150808-T2-02 572 IV19 Lin 6 P1 M2ERC0039-20150809-T1-01 386 IV20 Lin 8= D. mesozeirus P1 M2ERC0039-20150809-T1-01 IV21 Lin 8= D. mesozeirus P1 M2ERC027-t2-01 761 IV22 Lin 8= D. mesozeirus P1 M2ERC027-t2-01 IV23 Lin 8= D. mesozeirus P1 M2ERC049-t2-01 752 IV24 Lin 8= D. mesozeirus P1 M2ERC053-t1-01 772 IV25 Lin 8= D. mesozeirus P1 M2ERC053-t1-01 IV26 Lin 8= D. mesozeirus P1 M2ERC0053-T2-02 777 IV27 Lin 8= D. mesozeirus P1 M2ERC0053-T2-02 IV28 Lin 8= D. mesozeirus P1 MEARC3790.20150606-01 3 IV29 Lin 3 P1 MEARC3790.20150606-01 IV30 Lin 3 P1 MEARC3790.20150606-01 IV31 Lin 3 P1 MEARC3790.20150606-01 IV32 Lin 3 P1 MEARC3790-20150807-T1-01 409 IV33 Lin 3 P1 MEARC3811-20150807-T1-01 389 IV34 Lin 3 P1 MEARC4273-t1-01 756 IV35 Lin 4 P1 MEARC4400-t1-02 766 IV36 Lin 3 P1 MEBRC0027T2-01 673 IV37 Lin 1= P. bythius P1 MEBRC0027T2-01 IV38 Lin 1= P. bythius P1 MEBRC0027T3-01 675 IV39 Lin 1= P. bythius P1 MELUnk02-t2-01 757 IV40 Lin 4 P1 MELUnk02-t2-01 IV41 Lin 4 P1 MELUNK03-t1-01 758 IV42 Lin 4

13 Molecular systematics of subfauna from the Robe Valley

P1 MELUNK06-201508-T2-01 366 IV43 Lin 4 P1 MELUNK06-20150809-T1-01 372 IV44 Lin 4 P1 MELUNK07-20150809-T2-02 469 IV45 Lin 4 P1 MELUNK09T1-01 693 IV46 Lin 4 P1 Melunk10-t1-01 789 IV47 Lin 4 P1 Melunk10-t1-01 IV48 Lin 4 P1 RC13MEN0005-20150808-T1-01 472 IV49 Lin 4 P1 RC13MEN0005-20150808-T1-01 IV50 Lin 4 P1 RC13MRR0014-t1-01 788 IV51 Lin 9 P1 RC13MRR0026-t2-01 780 IV52 Lin 7 P1 RC13MRR0051T2-01 688 IV53 Lin 9 P1 RC13MRR0051T2-01 IV54 Lin 9 P1 RC13MRR0077-T1-02 717 IV55 Lin 9 P1 RC14MEB0071-01 751 IV56 Lin 1= P. bythius P1 RC14MEB0081-t2-01 771 IV57 Lin 1= P. bythius P1 RC14MEB0081-t2-01 IV58 Lin 1= P. bythius P2 2402EUNK05-20151002-T1-01 423 IV59 Lin 8= D. mesozeirus P2 2402EUNK11-20151002-T2-01 302 IV60 Lin 8= D. mesozeirus P2 DD14MEC0002-20150929-T3-03 508 IV61 Lin 1= P. bythius P2 DD14MEC0002-20150929-T3-03 IV62 Lin 1= P. bythius P2 DD14MEL0002-20151001-T2-01 646 IV63 Lin 4 P2 DD14MEL0002-20151001-T2-01 IV64 Lin 4 P2 DD14MEL0002-20151001-T2-01 IV65 Lin 4 P2 DD14MEM0001-20151001-T1-01 601 IV66 Lin 4 P2 DD14MEM0002-20151001-T2-01 611 IV67 Lin 4 P2 DD14MEM0003-20151001-T1-01 541 IV68 Lin 4 P2 DD14MRR0008-20151002-T3-01 449 IV69 Lin 6 P2 DD14MRR0009-20151002-T2-01 432 IV70 Lin 6 P2 DD14MRR0009-20151002-T3-03 439 IV71 Lin 6 P2 M2ERC0057-20151002-T2-01 407 IV72 Lin 8= D. mesozeirus P2 MEARC3790-20150930-T1-01 486 IV73 Lin 3 P2 MELUNK02-20151001-T1-01 624 IV74 Lin 4 P2 MELUNK02-20151001-T1-01 IV75 Lin 4 P2 MELUNK11-20151001-T2-01 653 IV76 Lin 4 P2 RC13MRR0026-20151002-T2-02 395 IV77 Lin 7 P2 RC14MEB0029-20151001-T1-02 133 IV78 Lin 1 P2 RC14MEB0060-20151001-T2-01 203 IV79 Lin 5 P2 RC14MEB0150-20151001-T2-01 38 IV80 Lin 1= P. bythius P2 RC14MRR0017-20151002-T2-01 380 IV81 Lin 10 P2 TOBRC0023-20151001-T1-01 31 IV82 Lin 3 P1 DD11MEC0003T2-01 685 IV83 Lin 1= P. bythius P1 DD11MEC0003T2-01 IV84 Lin 1= P. bythius P1 DD11MEC0003T3-01 686 IV85 Lin 1= P. bythius P1 DD11MEC0003T3-01 IV86 Lin 1= P. bythius P1 DD14MEC001-20150810-T1-01 559 IV87 Lin 1= P. bythius

14 Molecular systematics of subfauna from the Robe Valley

P1 MERC0114-T3-01 600 IV88 Lin 1= P. bythius P1 MERC0114-T2-04 604 P1 Nonumber-t1-01 792 IV89 Lin 4 P1 Nonumber-t1-01 IV90 Lin 4 P1 Nonumber-t2-01 793 IV91 Lin 4

Table 2. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Schizomida detected in the NJ analysis as shown in Figure 1.

lineage 1 2 3 4 4a 5 6 7 8 9 10 lin 1 0.007 0.008 0.010 0.009 0.012 0.013 0.013 0.011 0.012 0.010 lin 2 0.065 0.007 0.009 0.008 0.012 0.013 0.013 0.012 0.013 0.011 lin 3 0.080 0.058 0.009 0.010 0.011 0.013 0.013 0.012 0.011 0.010 lin 4 0.105 0.104 0.096 0.005 0.011 0.013 0.012 0.012 0.012 0.011 Lin 4a 0.113 0.097 0.099 0.031 0.011 0.013 0.012 0.012 0.013 0.011 lin 5 0.158 0.144 0.140 0.149 0.150 0.015 0.013 0.011 0.013 0.012 lin 6 0.161 0.149 0.141 0.147 0.151 0.169 0.010 0.012 0.012 0.011 lin 7 0.161 0.153 0.157 0.141 0.144 0.170 0.087 0.009 0.011 0.011 lin 8 0.155 0.141 0.142 0.131 0.135 0.157 0.081 0.065 0.012 0.010 lin 9 0.159 0.147 0.139 0.130 0.136 0.180 0.123 0.122 0.117 0.011 lin 10 0.150 0.143 0.139 0.148 0.151 0.172 0.145 0.155 0.148 0.167

Table 3. Mean distance (D) and standard error (s.e.) within lineages of Schizomida detected in the NJ analysis as shown in Figure 1 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis.

Lineage D s.e. N rep lin 1 0.007 0.002 18 IV61 lin 2 n/c n/c 1 IV07 lin 3 0.011 0.002 9 IV82 lin 4 0.008 0.001 26 IV76 lin 4a n/c n/c 1 IV09 lin 5 n/c n/c 1 IV79 lin 6 0.001 0.001 7 IV17 lin 7 0.002 0.001 6 IV14 lin 8 0.005 0.001 18 IV27 lin 9 0.013 0.004 4 IV55 lin 10 n/c n/c 1 IV81

Table 4. (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Schizomida detected at Robe Valley and the reference lineages as shown in Figure 2.

15 Molecular systematics of subfauna from the Robe Valley

Table 5. Distribution of the eight Schizomida lineages detected in the present study.

Lineage drillholes deposits 1 DD11MEB001, DD11MEC0003, Mesa B, Mesa DD14MEC001, DD14MEC002, C MEBRC0027, MERC0114, RC14MEB0029, RC14MEB0071, RC14MEB0081, RC14MEB0150 2 Budgie20150604 Waramboo 3 MEARC3790, MEARC3811, MEARC4400, Highway and TOBRC0023, Tod bore 4 DD14MEL0002, DD14MEM0001, Mesa L, Mesa DD14MEM0002, DD14MEM0003, M, Mesa N, DD14MEN0001, MEARC4273, Highway and MELUNK02, MELUNK03, MELUNK06, Tod bore MELUNK07, MELUNK09, MELUNK10, MELUNK11, nonumber, RC13MEN0005 4a DD14MEL0001 Mesa L 5 RC14MEB0060 Mesa B 6 DD14MRR007, DD14MRR008, 2402D DD14MRR009 7 DD14MRR004, DD14MRR005, 2402A DD14MRR0026, 8 M2ERC0027, M2ERC0039, M2ERC0049, 2402E M2ERC0053, M2ERC0057, 2402EUNK03, 2402EUNK05, 2402EUNK08, 2402EUNK09, 2402EUNK11, 2402EUNK13 9 RC13MRR0014, RC13MRR0051 2403D 10 RC14MRR0017 2401A

Table 6. Specimens of Amphipoda used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data

Phase Tracking number Cap no. Helix ID Lineage P1 Budgie.20150604-02 7 IV92 Lineage 2 P1 Budgie.20150604-02 IV93 Lineage 1 P1 Budgie.20150604-02 IV94 Lineage 8 P1 Budgie.20150604-02 IV95 Lineage 8 P1 Budgie.20150604-02 IV96 nd P1 Camp-20150604-02 15 IV97 nd P1 Camp-20150604-02 IV98 nd P1 Daisy-20150604-02 13 IV99 nd P1 Daves-20150604-01 1 IV100 Lineage 8 P1 Daves-20150604-01 IV101 nd P1 Daves-20150604-01 IV102 nd P1 Daves-20150604-01 IV103 nd P1 Daves-20150604-01 IV104 Lineage 8 P1 MB13WARR001-20150605-01 20 IV105 Lineage 5 P1 MB13WARR001-20150605-01 IV106 Lineage 5 P1 MB13WARR001-20150605-01 IV107 Lineage 5 P1 MB13WARR001-20150605-01 IV108 Lineage 5 P1 MB13WARR001-20150605-01 IV109 Lineage 5

16 Molecular systematics of subfauna from the Robe Valley

P1 MB13WARR010.20150606-01 18 IV110 nd P1 MB13WARR010.20150606-01 IV111 Lineage 3 P1 MB13WARR010.20150606-01 IV112 Lineage 3 P1 MB13WARR010.20150606-01 IV113 nd P1 MB13WARR010.20150606-01 IV114 Lineage 3 P1 TOBRC009-20150605-01 19 IV115 nd P1 TOBRC009-20150605-01 IV116 nd P1 TOBRC009-20150605-01 IV117 nd P1 TOBRC009-20150605-01 IV118 nd P1 TOBRC009-20150605-01 IV119 nd P1 Budgie-20150930-01 7 IV120 Lineage 1 P1 Budgie-20150930-01 IV121 Lineage 1 P1 Budgie-20150930-01 IV122 nd P1 Budgie-20150930-01 IV123 Lineage 1 P1 Budgie-20150930-01 IV124 nd P2 Camp-20150930-02 14 IV125 Lineage 1 P2 Camp-20150930-02 IV126 Lineage 1 P2 Daisy-201509-02 12 IV127 Lineage 2 P2 DAVES-20150930-04 4 IV128 Lineage 4 P2 DAVES-20150930-04 IV129 Lineage 7 P2 DAVES-20150930-04 IV130 Lineage 8 P2 DAVES-20150930-04 IV131 nd P2 DAVES-20150930-04 IV132 Lineage 4 P2 RC13MEA0279-20150930-02 17 IV133 nd P2 TOBRC0099-20150930-01 18 IV134 Lineage 6 P2 TOBRC0099-20150930-01 IV135 nd P2 TOBRC0099-20150930-01 IV136 nd P2 TOBRC0099-20150930-01 IV137 nd P2 TOBRC0099-20150930-01 IV138 nd

Table 7. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Amphipoda detected in the NJ analysis as shown in Figure 3.

Lineage lin 1 lin 2 lin 3 lin 4 lin 5 lin 6 lin 7 lin 8 lin 1 0.009 0.010 0.013 0.013 0.012 0.016 0.017 lin 2 0.067 0.009 0.013 0.011 0.013 0.016 0.018 lin 3 0.086 0.075 0.012 0.012 0.013 0.015 0.016 lin 4 0.099 0.107 0.107 0.011 0.012 0.017 0.017 lin 5 0.137 0.138 0.147 0.143 0.013 0.016 0.017 lin 6 0.145 0.145 0.148 0.149 0.156 0.017 0.017 lin 7 0.286 0.281 0.279 0.289 0.290 0.264 0.012 lin 8 0.304 0.300 0.298 0.321 0.300 0.298 0.151

Table 8. Mean distance (D) and standard error (s.e.) within lineages of Amphipoda detected in the NJ analysis as shown in Figure 3 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis.

17 Molecular systematics of subfauna from the Robe Valley

Lineage D s.e. N rep lin 1 0.011 0.003 6 IV126 lin 2 0.001 0.001 2 IV92 lin 3 0.001 0.001 3 IV111 lin 4 0.000 0.000 2 IV128 lin 5 0.018 0.003 5 IV108 lin 6 n/c n/c 1 IV134 lin 7 n/c n/c 1 IV129 lin 8 0.005 0.002 5 IV100

Table 9 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Amphipoda detected at Robe Valley and the reference lineages as shown in Figure 4.

Table 10. Distribution of the eight Amphipoda lineages detected in the present study.

Lineage drillholes deposits 1 Budgie, Camp Waramboo 2 Budgie, Daisy Waramboo 3 MB13WARR010 Waramboo 4 Daves Waramboo 5 MB13WARR001 Waramboo 6 TOBRC009 Highway and Tod bore 7 Daves Waramboo 8 Budgie, Daves Waramboo

Table 11. Specimens of Chilopoda used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking number Cap no. Helix ID Lineage P1 DD14MRR0004T1-01 694 IV139 Lineage 1 P1 DD14MRR0004T1-01 694 IV140 Lineage 1 P1 DD14MRR0005-T1-04 607 IV141 Lineage 1 P2 DD14MEL0001-20151001-T1-01 632 IV142 nd P2 MEARC4383-20151001-T2-01 217 IV143 Lineage 2

Table 12 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Chilopoda detected at Robe Valley and the reference lineages as shown in Figure 5.

Table 13. Distribution of the two Chilopoda lineages detected in the present study.

Lineage drillholes deposits 1 DD14MRR0004, DD14MRR0005 2304A 2 MEARC4383 Highway/Tod bore

Table 14. Specimens of Coleoptera used in the present study and the lineage to which they were assigned, based on variation at the COXI gene.

Phase Drillhole Cap no. Helix ID Lineage P1 DD14MRR0004T1-03 696 IV144 Lineage 2 P1 DD14MRR0004T1-03 IV145 Lineage 2

18 Molecular systematics of subfauna from the Robe Valley

P1 DD14MRR0004T1-03 IV146 Lineage 2 P1 DD14MRR0004T1-03 IV147 Lineage 2 P1 DD14MRR009-20150808-T3-01 562 IV148 Lineage 6 P1 RC14MEB0068-20150807-T1-02 424 IV149 nd P2 RC14MEB0060-20151001-T2-02 204 IV150 Lineage 4 P1 MEARC3814-20150807-T1-02 379 IV151 Lineage 1 P1 MEARC4400-t1-01 765 IV152 Lineage 5 P1 MEARC4400-t1-01 IV153 Lineage 3

Table 15 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Coleoptera detected at Robe Valley and the reference lineages as shown in Figure 6.

Table 16. Distribution of the six Coleoptera lineages detected in the present study.

Lineage drillholes deposits 1 MEARC3814 Highway/Tod bore 2 DD14MRR0004 2403A 3 MEARC4400 Highway/Tod bore 4 RC14MEB0060 Mesa B 5 MEARC4400 Highway/Tod bore 6 DD14MRR0009 2402D

Table 17. Specimens of Polydesmida (Diplopoda) used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data. Phase Tracking number Cap no. Helix ID Lineage P1 MEARC2401-20150807-T2-02 400 IV190 nd P1 MEARC3500T2-02 691 IV191 nd P1 RC13MEA0279-20150807-T1-01 382 IV192 nd P1 RC13MEA0279-20150807-T1-01 382 IV193 nd P1 RC13MEA0279-20150807-T1-01 382 IV194 nd P2 DD14MEB0005-20151001-T2-02 81 IV195 nd P2 DD14MEB0005-20151001-T3-01 84 IV196 nd P2 DD14MEB0005-20151001-T3-01 84 IV197 nd P2 DD14MEB0005-20151001-T3-01 84 IV198 nd P2 MEARC3500-20150930-T1-02 267 IV199 nd P2 MEARC3500-20150930-T1-02 267 IV200 nd

Table 18. Specimens of Diplura used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking number Cap no. Helix ID Lineage P1 DD11MEB001T1-01 679 IV154 Lineage 1 P1 DD11MEB001T1-01 IV155 Lineage 2 P1 DD11MEC0005-02 7 IV156 Lineage 4 P2 DD14MEB0005-20151001-T3-02 85 IV157 Lineage 3 P2 MEARC5017-20151001-T2-01 65 IV158 nd P2 RC14MEB0101-20151001-T2-01 93 IV159 Lineage 3 P1 MEARC3790-20150807-T2-03 414 IV160 nd

19 Molecular systematics of subfauna from the Robe Valley

Table 19 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Diplura detected at Robe Valley and the reference lineages as shown in Figure 7.

Table 20. Distribution of the four Diplura lineages detected in the present study.

Lineage drillholes deposits 1 DD11MEB001 Mesa B 2 DD11MEB001 Mesa B 3 DD14MEB0005, RC14MEB0101 Mesa B 4 DD11MEC0005 Mesa C

Table 21. Specimens of Isopoda used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking number Cap no. Helix ID Lineage P1 M2ERC027-t2-02 762 IV163 Lineage 1 P1 M2ERC029-t2-01 769 IV164 nd P1 M2ERC053-T2-01 776 IV165 Lineage 1 P1 M2ERC053-T2-01 IV166 nd P1 M2ERC053-T2-01 IV167 Lineage 1 P1 M2ERC053-T2-01 IV168 nd P1 RC13MRR0026-t2-02 781 IV169 nd P2 M2ERC0057-20151002-T2-03 409 IV170 nd P2 MEARC4923-20150930-T1-01 343 IV171 Lineage 9a P2 MELUNK02-20151001-T1-03 626 IV172 Lineage 3 P2 MELUNK02-20151001-T2-02 629 IV173 Lineage 3 P2 MELUNK02-20151001-T2-02 IV174 Lineage 3 P2 RC13MRR0014-20151002-T1-01 548 IV175 Lineage 2 P2 RC14MEB0060-20151001-T2-03 205 IV176 Lineage 5 P2 RC14MEB0088-20151001-T3-01 576 IV177 Lineage 8 P2 RC14MEB0101-20151001-T1-01 89 IV178 Lineage 4 P2 TOBRC0020-20151001-T2-02 13 IV179 Lineage 7 P2 TOBRC0020-20151001-T2-02 IV180 nd P1 MEARC2401-20150807-T2-03 401 IV181 Lineage 9b P1 MEARC3811-20150807-T1-04 392 IV182 nd P1 MEARC4795-20150807-T1-03 269 IV183 nd P1 MEARC4795-20150807-T2-02 273 IV184 nd P1 MEARC4795-20150807-T2-02 IV185 nd P1 MEARC4795-20150807-T2-02 IV186 nd P1 MEARC4795-20150807-T2-02 IV187 nd P1 MEARC5053T3-03 541 IV188 Lineage 9b P1 TOBRC0020-t1-01 784 IV189 Lineage 6

20 Molecular systematics of subfauna from the Robe Valley

Table 22. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Isopoda detected in the NJ analysis as shown in Figure 8.

Lineage 1 2 3 4 5 6 7 8 9a 9b 1 0.011 0.016 0.017 0.016 0.016 0.016 0.016 0.016 0.014 2 0.070 0.016 0.015 0.018 0.016 0.017 0.015 0.016 0.014 3 0.201 0.191 0.017 0.017 0.016 0.015 0.015 0.015 0.015 4 0.189 0.191 0.194 0.017 0.016 0.015 0.017 0.015 0.015 5 0.244 0.242 0.259 0.224 0.010 0.010 0.016 0.017 0.017 6 0.239 0.236 0.242 0.235 0.076 0.009 0.016 0.016 0.015 7 0.238 0.232 0.238 0.227 0.082 0.067 0.016 0.017 0.016 8 0.228 0.220 0.226 0.238 0.218 0.211 0.207 0.014 0.014 9a 0.227 0.224 0.238 0.201 0.231 0.216 0.201 0.166 0.006 9b 0.231 0.225 0.230 0.204 0.223 0.207 0.204 0.167 0.037

Table 23. Mean distance (D) and standard error (s.e.) within lineages of Isopoda detected in the NJ analysis as shown in Figure 8 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis. Lineage D s.e. N Rep 1 0.002 0.001 3 IV167 2 n/c n/c 1 IV175 3 0.001 0.001 3 IV172 4 n/c n/c 1 IV178 5 n/c n/c 1 IV176 6 n/c n/c 1 IV189 7 n/c n/c 1 IV179 8 n/c n/c 1 IV177 9a n/c n/c 1 IV171 9b 0.025 0.006 2 IV181

Table 24 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Isopoda detected at Robe Valley and the reference lineages as shown in Figure 9.

Table 25. Distribution of the ten Isopoda lineages detected in the present study.

Lineage drillholes deposits 1 M2ERC027 2402E 2 RC13MRR0014 2403A,B,D 3 MELUNK02 Mesa L 4 RC14MEB0101 Mesa B 5 RC14MEB0060 Mesa B 6 TOBRC0020 Highway/Tod bore 7 TOBRC0020 Highway/Tod bore 8 RC14MEB0088 Mesa B 9a MEARC4923 Highway/Tod bore 9b MEARC2401, MEARC5053 Highway/Tod bore

21 Molecular systematics of subfauna from the Robe Valley

Table 26. Specimens of Pseudoscorpiones used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking number Cap. No. Helix ID Lineage P1 DD14MEB0002-20150607-01sc 5 IV201 Hyiidae 1 P1 DD14MRR0004T1-04 697 IV202 Hyiidae 5 P1 DD14MRR0004T1-04 IV203 Chthoniidae 11 P1 DD14MRR0005-T1-05 608 IV204 Chthoniidae 11 P1 DD14MRR0005-T2-01 773 IV205 Atemnidae? 13 P1 DD14MRR0008-20150808-T1-02 484 IV206 Hyiidae 3 P1 DD14MRR0008-20150808-T2-03 491 IV207 Atemnidae? 15 P1 M2ERC027-t2-03 763 IV208 Atemnidae? 14 P1 M2ERC027-t2-03 IV209 Hyiidae 4 P1 MEARC3814-20150606-02sc 2 IV210 Chthoniidae 12 P1 RC13MEA0279-20150807-T1-02 383 IV211 Hyiidae 6 P2 DD14MEL0001-20151001-T2-02 637 IV212 Chthoniidae 9 P2 DD14MRR0009-20151002-T2-02 433 IV213 Hyiidae 7 P2 DD14MRR0009-20151002-T3-01 437 IV214 Atemnidae? 15 P2 MEARC3814-20150930-T1-01 279 IV215 Chthoniidae 12 P2 RC14MEB0010-20151001-T3-01 122 IV216 nd P2 RC14MEB0029-20151001-T1-01 132 IV217 Hyiidae 2 P2 RC14MEB0060-20151001-T3-02 209 IV218 Hyiidae 8 P2 RC14MEB0101-20151001-T1-02 90 IV219 nd P2 RC14MEB0115-20151001-T1-01 101 IV220 nd P2 RC14MEB0115-20151001-T1-01 IV221 nd P2 RC14MEB0123-20151001-T2-01 310 IV222 nd P2 RC14MEB0123-20151001-T2-01 IV223 nd P2 RC14MEB0123-20151001-T3-01 313 IV224 nd P2 RC14MEB0123-20151001-T3-01 IV225 nd P2 RC14MRR0020-20151002-T2-01 369 IV226 Chthoniidae 10

22 Molecular systematics of subfauna from the Robe Valley

Table 27. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Pseudoscorpiones detected in the NJ analysis as shown in Figure 10. Distances between lineages within each family are highlighted in yellow.

Hyiidae Chthoniidae Atemnidae Lineage 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 1 0.008 0.015 0.015 0.017 0.016 0.016 0.016 0.018 0.018 0.016 0.018 0.016 0.015 0.014 2 0.051 0.016 0.016 0.017 0.016 0.015 0.016 0.019 0.020 0.016 0.017 0.017 0.015 0.015 3 0.221 0.227 0.007 0.016 0.015 0.017 0.016 0.017 0.017 0.017 0.016 0.017 0.016 0.017 4 0.211 0.218 0.038 0.017 0.015 0.017 0.016 0.017 0.017 0.017 0.016 0.017 0.016 0.017 5 0.265 0.262 0.265 0.262 0.015 0.016 0.016 0.016 0.015 0.017 0.016 0.015 0.014 0.015 6 0.232 0.227 0.251 0.245 0.240 0.012 0.013 0.016 0.017 0.016 0.015 0.015 0.013 0.014 7 0.254 0.254 0.264 0.249 0.240 0.133 0.012 0.015 0.016 0.015 0.014 0.015 0.016 0.015 8 0.251 0.245 0.272 0.258 0.244 0.150 0.133 0.015 0.016 0.015 0.015 0.014 0.013 0.013 9 0.275 0.274 0.284 0.287 0.268 0.255 0.257 0.263 0.012 0.015 0.016 0.016 0.016 0.016 10 0.265 0.265 0.283 0.286 0.266 0.258 0.252 0.257 0.105 0.015 0.016 0.019 0.017 0.017 11 0.264 0.274 0.277 0.272 0.274 0.277 0.254 0.267 0.194 0.204 0.014 0.016 0.014 0.015 12 0.274 0.278 0.257 0.251 0.283 0.265 0.247 0.266 0.210 0.226 0.165 0.016 0.014 0.014 13 0.290 0.295 0.293 0.283 0.280 0.284 0.288 0.285 0.288 0.286 0.291 0.286 0.013 0.012 14 0.258 0.267 0.273 0.261 0.274 0.257 0.254 0.251 0.281 0.271 0.264 0.266 0.119 0.009 15 0.273 0.286 0.299 0.282 0.292 0.273 0.263 0.275 0.288 0.294 0.282 0.285 0.146 0.076

23 Molecular systematics of subfauna from the Robe Valley

Table 28. Mean distance (D) and standard error (s.e.) within lineages of Pseudoscorpiones detected in the NJ analysis as shown in Figure 10 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis. Lineage D s.e. N Ref 1 n/c n/c 1 IV201 2 n/c n/c 1 IV217 3 n/c n/c 1 IV206 4 n/c n/c 1 IV209 5 n/c n/c 1 IV202 6 n/c n/c 1 IV211 7 n/c n/c 1 IV213 8 n/c n/c 1 IV219 9 n/c n/c 1 IV212 10 n/c n/c 1 IV226 11 0.001 0.001 2 IV203 12 0.001 0.001 2 IV210 13 n/c n/c 1 IV205 14 n/c n/c 1 IV208 15 0.012 0.004 2 IV207

Table 29 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Chthoniidae (Pseudoscorpiones) detected at Robe Valley and the reference lineages as shown in Figure 11.

Table 30 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Hyiidae and Atemnidae (Pseudoscorpiones) detected at Robe Valley and the reference lineages as shown in Figure 12.

Table 31. Distribution of the 15 Pseudoscorpiones lineages detected in the present study.

Lineage drillholes deposits Hyiidae 1 DD14MEB0002 Mesa B 2 RC14MEB0029 Mesa B 3 DD14MRR0008 2402D 4 M2ERC027 2402E 5 DD14MRR0004 2403A 6 RC13MEA0279 Highway/Tod bore 7 DD14MRR0009 2402D 8 RC14MEB0101 Mesa B Chthoniidae 9 DD14MEL0001 Mesa L 10 RC14MRR020 2401A 11 DD14MRR0004, DD14MRR0005 2403A 12 MEARC3814 Highway/Tod bore Atemnidae 13 DD14MRR0005 2403A 14 M2ERC027 2402E 15 DD14MRR0008, DD14MRR0009 2402D

24 Molecular systematics of subfauna from the Robe Valley

Table 32. Specimens of Thysanura used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data.

Phase Tracking number Cap. No. Helix ID Lineage P2 MEARC4383-20151001-T1-01 213 IV227 nd P2 RC14MEB0060-20151001-T3-01 208 IV228 nd P2 RC14MEB0060-20151001-T3-01 IV229 nd P1 MEARC3814-20150606-01sc 1 IV230 nd

25 Molecular systematics of subfauna from the Robe Valley

IV63 DD14MEL0002-20151001-T2-01 Schizomida 61 IV65 DD14MEL0002-20151001-T2-01 Schizomida Figure 1. Neighbour-joining IV43 MELUNK06-201508-T2-01 Schizomida 20 IV45 MELUNK07-20150809-T2-02 Schizomida analysis of specimens of 19 IV90 Nonumber-t1-01 Schizomida Schizomida from the present IV35 MEARC4273-t1-01 Schizomida study. Numbers on major nodes 3592 IV41 MELUnk02-t2-01 Schizomida 60 IV75 MELUNK02-20151001-T1-01 Schizomida correspond to bootstrap IV47 Melunk10-t1-01 Schizomida IV42 MELUNK03-t1-01 Schizomida support over 100 iterations; 19 59 IV10 DD14MEM0001T2-03 Schizomida values <50% are not shown. 61 IV68 DD14MEM0003-20151001-T1-01 Schizomida 39 IV64 DD14MEL0002-20151001-T2-01 Schizomida Scale bar= genetic distance. lin 4 IV48 Melunk10-t1-01 Schizomida The specimens used to IV76 MELUNK11-20151001-T2-01 Schizomida 24 IV74 MELUNK02-20151001-T1-01 Schizomida represent each lineage in the IV89 Nonumber-t1-01 Schizomida model-based phylogenetic IV91 Nonumber-t2-01 Schizomida 36 IV44 MELUNK06-20150809-T1-01 Schizomida analysis are highlighted in 6 IV40 MELUnk02-t2-01 Schizomida yellow. 9834 IV46 MELUNK09T1-01 Schizomida IV67 DD14MEM0002-20151001-T2-01 Schizomida 72 IV66 DD14MEM0001-20151001-T1-01 Schizomida IV49 RC13MEN0005-20150808-T1-01 Schizomida 100 100 IV11 DD14MEN0001-20150808-T2-02 Schizomida 81 IV50 RC13MEN0005-20150808-T1-01 Schizomida IV09 DD14MEL0001-t2-01 Schizomida lin 4a 18 IV29 MEARC3790.20150606-01 Schizomida 16 IV34 MEARC3811-20150807-T1-01 Schizomida 20 IV33 MEARC3790-20150807-T1-01 Schizomida 73 93 IV31 MEARC3790.20150606-01 Schizomida 100 IV30 MEARC3790.20150606-01 Schizomida lin 3 82 IV32 MEARC3790.20150606-01 Schizomida 92 IV73 MEARC3790-20150930-T1-01 Schizomida 97 IV36 MEARC4400-t1-02 Schizomida IV82 TOBRC0023-20151001-T1-01 Schizomida IV07 Budgie20150604-06 Schizomida lin 2 24 IV3 9 MEBRC0027T3-01 Schizomida 100 45 IV58 RC14MEB0081-t2-01 Schizomida 39 IV57 RC14MEB0081-t2-01 Schizomida 34 IV37 MEBRC0027T2-01 Schizomida 48 49 IV78 RC14MEB0029-20151001-T1-02 Schizomida 85 IV38 MEBRC0027T2-01 Schizomida 82 IV80 RC14MEB0150-20151001-T2-01 Schizomida IV56 RC14MEB0071-01 Schizomida lin 1 100 IV62 DD14MEC0002-20150929-T3-03 Schizomida IV83 DD11MEC0003T2-01 Schizomida 100 27 IV61 DD14MEC0002-20150929-T3-03 Schizomida 34 IV84 DD11MEC0003T2-01 Schizomida 73 IV86 DD11MEC0003T3-01 Schizomida IV85 DD11MEC0003T3-01 Schizomida 66 IV87 DD14MEC001-20150810-T1-01 Schizomida 59 IV88 MERC0114-T3-01 Schizomida IV81 RC14MRR0017-20151002-T2-01 Schizomida lin 10 85 IV51 RC13MRR0014-t1-01 Schizomida IV54 RC13MRR0051T2-01 Schizomida 100 lin 9 IV53 RC13MRR0051T2-01 Schizomida 89 IV55 RC13MRR0077-T1-02 Schizomida 52 74 IV17 DD14MRR007-20150808-T1-02 Schizomida 41 IV18 DD14MRR0008-20150808-T3-01 Schizomida 25 IV16 DD14MRR007-20150808-T2-01 Schizomida 17 IV71 DD14MRR0009-20151002-T3-03 Schizomida lin 6 64 IV70 DD14MRR0009-20151002-T2-01 Schizomida 91 100 IV69 DD14MRR0008-20151002-T3-01 Schizomida IV19 DD14MRR009-20150808-T2-02 Schizomida 37 IV14 DD14MRR0005-T1-01 Schizomida 48 IV52 RC13MRR0026-t2-01 Schizomida IV15 DD14MRR0005-T2-03 Schizomida 57 lin 7 60 IV77 RC13MRR0026-20151002-T2-02 Schizomida 100 100 IV13 DD14MRR0005-T1-01 Schizomida IV12 DD14MRR0004-t2-01 Schizomida 91 IV27 M2ERC0053-T2-02 Schizomida 73 IV28 M2ERC0053-T2-02 Schizomida 60 IV06 2402EUNK13T2-01 Schizomida 19 IV03 2402eUnk08-20150809-T1-01 Schizomida 89 4 IV60 2402EUNK11-20151002-T2-01 Schizomida IV01 2402eUnk03-20150809-T1-01 Schizomida 20 49 IV21 M2ERC0039-20150809-T1-01 Schizomida IV04 2402eUNK09-T3-01 Schizomida 57 IV23 M2ERC027-t2-01 Schizomida lin 8 100 IV22 M2ERC027-t2-01 Schizomida 15 IV24 M2ERC049-t2-01 Schizomida 21 IV26 M2ERC053-t1-01 Schizomida 24 IV20 M2ERC0039-20150809-T1-01 Schizomida 18 IV72 M2ERC0057-20151002-T2-01 Schizomida 3 IV59 2402EUNK05-20151002-T1-01 Schizomida IV05 2402eUnk11-20150809-T2-01 Schizomida 3 19 IV02 2402EUNK05T3-01 Schizomida 9 IV25 M2ERC053-t1-01 Schizomida IV79 RC14MEB0060-20151001-T2-01 Schizomida lin 5

0.02 26 Molecular systematics of subfauna from the Robe Valley

Figure 2. Bayesian analysis of COXI haplotypes of Schizomida from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site. EU272675 Brignolozomus woodwardi SzA

IV61 DD14MEC0002 20150929 T3 03 lin 1 1.00 0.55 Paradraculoides bythius WAM T63344 0.81 IV07 Budgie20150604 06 lin 2

1.00 Paradraculoides anachoretus WAM T66236 IV82 TOBRC0023 20151001 T1 01 lin 3 0.97 1.00 Paradraculoides sp OFB 2007 0.50 Paradraculoides gnophicola WAM T63371 3 1.00 Paradraculoides kryptus WAM T65802

1.00 IV76 MELUNK11 20151001 T2 01 lin 4 IV09 DD14MEL0001 t2 01 lin 4a C119 ex T95543 0.911.00 1.00 GH7 Draculoides Dra PES16433 GH8 Draculoides Dra PES16447 C37 T92541B 0.98 HE17 BHRC151 20141216 T3 01c 0.980.87 0.61 HE4 BHRC001 20141028 SC01 0.79 IS33 Kbrc116720150809t2 05 Schizomida sp IS15 KBRC1389 20150909 04 Schizomida 0.900.97 0.99 IT67 BHRC401 151013 T1 01 Schizomida C58 T98320 0.96 T93233 C17 CBRC099P5T1 3 Paradraculoides sp cardo IV17 DD14MRR007 20150808 T1 02 lin 6 1.00 IV14 DD14MRR0005 T1 01 lin 7 1.00 IV27 M2ERC0053 T2 02 lin 8 1.00 1.00 1.00 EU272730 Draculoides mesozeirus 1.00 IV55 RC13MRR0077 T1 02 lin 9 U72 Schizomid T95463 1.00 U74 Schizomid T95462 0.98 1.00 U83 Schizomid T95450 EU272684 Draculoides bramstokeri IV81 RC14MRR0017 20151002 T2 01 lin 10

1.00 FT4 Draculoides SCH012 PSB0166R 0.93 0.50 1.00 FT6 Draculoides SCH012 GBP553 1.00 GG9 Draculoides sp PSA0806R 1.00 FT5 Draculoides sp PSB0163R FZ21 Schizomida RC13KOOD0308P7T1 1 IV79 RC14MEB0060 20151001 T2 01 lin 5 0.1

27 Molecular systematics of subfauna from the Robe Valley

Figure 3. Neighbour-joining analysis of specimens of Amphipoda from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

IV93 Budgie.20150604-02 Amphipoda 65 IV120 Budgie-20150930-01 Amphipoda 100 IV123 Budgie-20150930-01 Amphipoda 100 lin 1 99 IV126 Camp-20150930-02 Amphipoda

IV121 Budgie-20150930-01 Amphipoda 66 100 IV125 Camp-20150930-02 Amphipoda

IV92 Budgie.20150604-02 Amphipoda 87 lin 2 100 IV127 Daisy-201509-02 Amphipoda

IV114 MB13WARR010.20150606-01 Amphipoda

100 IV111 MB13WARR010.20150606-01 Amphipoda lin 3 100 72 IV112 MB13WARR010.20150606-01 Amphipoda

IV128 DAVES-20150930-04 Amphipoda 93 lin 4 100 IV132 DAVES-20150930-04 Amphipoda

IV109 MB13WARR001-20150605-01 Amphipoda

IV105 MB13WARR001-20150605-01 Amphipoda 100 100 100 IV108 MB13WARR001-20150605-01 Amphipoda lin 5

87 IV106 MB13WARR001-20150605-01 Amphipoda

69 IV107 MB13WARR001-20150605-01 Amphipoda

IV134 TOBRC0099-20150930-01 Amphipoda lin 6

IV129 DAVES-20150930-04 Amphipoda lin 7

IV95 Budgie.20150604-02 Amphipoda

IV100 Daves-20150604-01 Amphipoda 100 IV130 DAVES-20150930-04 Amphipoda lin 8 89 IV94 Budgie.20150604-02 Amphipoda 12 47 IV104 Daves-20150604-01 Amphipoda

0.05

28 Molecular systematics of subfauna from the Robe Valley

Figure 4. Bayesian analysis of COXI haplotypes of Amphipoda from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

JX150976 Daphnia pulex voucher NZPL873 JN233925 Daphnia pulicaria voucher 0833 IV126 Camp 20150930 02 Amphipoda lin 1 1.00 0.94 rr3a 2b 1.00 IT22 BHRC231.20151014 03 Amphipoda IT06 BHRC118.20151013 02 Amphipoda

1.00 IV92 Budgie.20150604 02 Amphipoda lin 2 1.00 nr5miwell d

0.97 IV111 MB13WARR010.20150606 01 Amphipoda lin 3 0.77 GY19 BIL4nr 20141109 20 1.001.00 0.96 L8 107b Nedsia sp 2 IS59 KBRC1310 20150911 02 Amphipoda 1.00 EU304458 Nedsia sp. Caves Creek IV128 DAVES 20150930 04 Amphipoda lin 4 1.00 IV108 MB13WARR001 20150605 01 Amphipoda lin 5 1.00 Melitidae wc17 1 Nedsia douglasi 1.00 0.98 B3 60a Nedsia sp 1

1.00 IS43 RHWB006 20150910 01 Amphipoda 0.84 cob12 5a 1.00 pt03 11a IV134 TOBRC0099 20150930 01 Amphipoda lin 6 0.72 0.681.00 IS37 RHWB005 20150910 01 Amphipoda 0.79 IS38 RHWB005 20150910 01 Amphipoda pm01a 10 1.00 DH1 BIL4nr 1112 03 01 Amphipoda 1.00 1.00 L26 S5 200 unknown sp 3 BF14 X62P2 2011 01 Amphipoda

1.00 JQ608487 Norcapensis mandibulis IV129 DAVES 20150930 04 Amphipoda lin 7 0.97 IV100 Daves 20150604 01 Amphipoda lin 8 1.00 Niphargidae KC315635 Niphargus fontanus isolate UK HT3 1.00 KC315646 Niphargus glenniei isolate HT4 0.1

29 Molecular systematics of subfauna from the Robe Valley

Figure 5. Bayesian analysis of COXI haplotypes of Chilopoda from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

NC014273 Opisthopatus cinctipes EF624055 Metaperipatus inae IS87 KBRC136220150911T2 03 Geophilida sp 1.00 AB610774 Mecistocephalus multidentatus 0.73 AF334321 Paralamyctes monteithi Henicopidae

1.00 IV139 DD14MRR0004T1 01 Chilopoda 1.00 IV140 DD14MRR0004T1 01 Chilopoda Lineage 1 1.00 1.00 Geophilomorpha IV141 DD14MRR0005 T1 04 Chilopoda KF569297 Gnathoribautia bonensis 1.00 JN306685 Geophilus flavus 1.00 KR736251 Stenotaenia linearis AK10 PE111022 Cryptops DC10

0.99 BX11 PES 0968 Cryptops pilbara1 1.00 1.00 EY5 Cryptops sp B35 BX12 PES 3783 Cryptops pilbara2 1.00 0.83 CQ6 Cryptops sp. B30 1106 13 EX13 BX13 991 5262 Cryptops MH1 0.81 1.00 1.00 BX8 PES 5889 Cryptops MH2 1.00 0.57 CQ3 Cryptops sp. B32 CC1798 Cryptops 1.00 BX9 PES 5272 Cryptops MH2 BQ12 PES 5087 Cryptops won1 0.54 0.85 G349 Cryptopidae sp MJ LN9808 0.88 G124 Cryptops Upper South Fortescue

1.00 0.98 CQ5 nr Cryptops sp. B11 PCRC088 CQ2 nr Cryptops sp. B15 CCWUNK05 0.81 IV143 MEARC4383 20151001 T2 01 Chilopoda Lineage 2 AY288745 Scolopocryptops sexspinosus Cryptopidae

G123 Cryptops Upper South Fortescue 0.1

30 Molecular systematics of subfauna from the Robe Valley

Figure 6. Bayesian analysis of COXI haplotypes of Coleoptera from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

GU671532 Hemiptera sp. FJ456942 L. delicatula

IV144 DD14MRR0004T1 03 Coleoptera IV145 DD14MRR0004T1 03 Coleoptera lin 2 1.00 IV146 DD14MRR0004T1 03 Coleoptera IV147 DD14MRR0004T1 03 Coleoptera 0.94 IV153 MEARC4400 t1 01 Coleoptera lin 3 1.00 0.96 KJ962132 Stricticollis tobias Anthicidae KJ961916 Necrobia violacea voucher ZMUO Cleridae 0.87 IV151 MEARC3814 20150807 T1 02 Coleoptera lin 1 1.00 1.00 EU710801 Pterostichus mutus Carabidae HM180603 Harpalus discrepans Carabidae 0.85 BX6 PES 5290 Bembidion MH1 Carabidae 1.00 1.00 BX7 PES 5285 Bembidion MH1 Carabidae 0.86 Carabidae G344 LN9362 Anillini sp OP Carabidae PSF0019R LN6326 J11 1 Zuphiinae S1 Carabidae 1.00 0.93 CF17 PSC0889R Zuphiini sp B03 Carabidae 1.00 G348 LN8860 Zuphiini sp UM Carabidae CF18 edited JIN0968R Zuphiini sp. Carabidae 0.90 IV148 DD14MRR009 20150808 T3 01 Coleoptera lin 6 CF5 GB0002R Curculionidae Genus 1 sp. B06 0.50 CF6 GF0421R Curculionidae

1.000.80 CF9 SF0034R Curculionidae Genus 1 sp. B07 0.78 0.81 CF10 SF0139R Curculionidae Genus 1 sp. B07 0.57 CF13 SF0725R Curculionidae Genus 1 sp. B09 0.93 CF7 PSC0890R Curculionidae Curculionidae 1.00 CF8 PSD0107R Curculionidae Genus 1 sp. B08 IV150 RC14MEB0060 20151001 T2 02 Coleoptera lin 4 1.00 0.87 IV152 MEARC4400 t1 01 Coleoptera lin 5 1.00 HE615891 Trigonopterus sp. Curculionidae GU213687 Echinodera ibleiensis Curculionidae

0.1

31 Molecular systematics of subfauna from the Robe Valley

Figure 7. Bayesian analysis of COXI haplotypes of Diplura from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

Lycorma delicatula FJ456942 Hemiptera sp. GU671532 IV154 DD11MEB001T1 01 Diplura lin 1 1.00 IV155 DD11MEB001T1 01 Diplura lin 2 1.00 IV157 DD14MEB0005 20151001 T3 02 Diplura 0.97 1.00 lin 3 Campodeidae IV159 RC14MEB0101 20151001 T2 01 Diplura 0.99 AF370844 Campodea tillyardi 1.00 HQ882832 Lepidocampa weberi HQ943342 Diplura sp.

IV156 DD11MEC0005 02 Diplura lin 4 BX29 PES 5141 Parajapygidae MH1 1.00 1.00 Parajapygidae 0.95 BX30 PES 5190 Parajapygidae MH1 1.00 G331 LN9880 Parajapygidae sp MJ 0.99 G332 LN8894 Parajapygidae sp MJ 1.00 IT74 BHRC0514 151013 T2 02 Diplura 0.61 AY771989 Japyx solifugus 0.93 G330 LN8732 Japygidae sp MJ BX16 PES 5195 Japygidae MH1 0.90 0.85 BX17 PES 5952 Japygidae MH1 0.59 1.00 Japygidae 0.98 G432 110953 Japygidae sp Central Pilbara 1.00 0.96 G433 110848 Japygidae Central Pilbara sp BQ8 PES 4283 Japygidae won1 1.00 G329 LN9310 Japygidae sp MA G473 111138b Anajapygidae sp NS Anajapygidae BX40 PES 5878 Projapygidae MH1 0.78 BQ11 PES 5439 Projapygidae won1 BQ7 PES 4269 Projapygidae 1.00 1.00 Projapygidae 1.00 BQ10 PES 2546 Projapygidae

0.78 G336 LN9557 Projapygidae sp OP G333 LN8949 Projapygidae sp MJ 1.00 G334 LN9141 Projapygidae sp UM 0.1

32 Molecular systematics of subfauna from the Robe Valley

Figure 8. Neighbour-joining analysis of specimens of Isopoda from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

98 IV163 M2ERC027-t2-02 Isopoda 100 IV167 M2ERC053-T2-01 Isopoda lin 1 100 IV165 M2ERC053-T2-01 Isopoda

50 IV175 RC13MRR0014-20151002-T1-01 Isopoda lin 2 IV173 MELUNK02-20151001-T2-02 Isopoda 100 lin 3 100 IV172 MELUNK02-20151001-T1-03 Isopoda 76 IV174 MELUNK02-20151001-T2-02 Isopoda IV178 RC14MEB0101-20151001-T1-01 Isopoda lin 4

64 IV176 RC14MEB0060-20151001-T2-03 Isopoda lin 5 100 IV189 TOBRC0020-t1-01 Isopoda lin 6 IV179 TOBRC0020-20151001-T2-02 Isopoda lin 7 IV177 RC14MEB0088-20151001-T3-01 Isopoda lin 8 IV171 MEARC4923-20150930-T1-01 Isopoda lin 9a 94 IV181 MEARC2401-20150807-T2-03 Isopoda 100 lin 9b 89 IV188 MEARC5053T3-03 Isopoda

0.02

33 Molecular systematics of subfauna from the Robe Valley

Figure 9. Bayesian analysis of COXI haplotypes of Isopoda from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

DQ838028 Chydaekata acuminata DQ679986 Chydaekata sp. lin 1 1.00 IV167 M2ERC053 T2 01 Isopoda 1.00 IV175 RC13MRR0014 20151002 T1 01 Isopoda lin 2 1.00 IT40 BHRCOPP1 151013 T3 02 Isopoda BH lin 1 0.67 IS126 KBRC1154 20150909 T2 01 RH Lin 6 IV172 MELUNK02 20151001 T1 03 Isopoda lin 3 0.68 0.75 IV178 RC14MEB0101 20151001 T1 01 Isopoda lin 4

0.75 EU364629 Laevophiloscia yalgoorensis IS97 CWRC485.20150709 T1 04 RH Lin 5 IV171 MEARC4923 20150930 T1 01 Isopoda lin 9a 1.00 lin 9b 1.00 IV181 MEARC2401 20150807 T2 03 Isopoda 1.00 IE45 MEARC2999P6T2 1 Lin Mesa A- P6 IV177 RC14MEB0088 20151001 T3 01 Isopoda lin 8 EO11 Troglarmadillo ISO005 EKP0042 1.00 GA0136R J17 5 0.72 0.73 1.00 1.00 GFR002 J17 9 GSR0020 J17 11 0.58 IE37 BHRC122 20141216 T2 03 0.61 IE39 TBRC045P3T1 3 0.56 0.58 1.00 IS111 CWRC937 20150912 T2 01 RH Lin 2 IS92 RHRC414 20150709 T2 02 RH Lin 1

0.99 G92 Troglarmadillo sp LN8501 IS105 TSOOPP03 20150709 T2 03 RH Lin 3

1.00 IV176 RC14MEB0060 20151001 T2 03 Isopoda lin 5 1.00 IV189 TOBRC0020 t1 01 Isopoda lin 6 1.00 IV179 TOBRC0020 20151001 T2 02 Isopoda lin 7 1.00 0.95 EO12 Troglarmadillo sp. B42 EEX0561 1.00 G350 LN9740 Troglamadillo sp OP 1.00 EXR1356 J17 2 1.00 0.99 EO10 Troglarmadillo sp. B39 MG0198R IE40 CWRC155P3T2 1 0.73 AB626254 Burmoniscus sp. FN824099 Armadillidium nasatum 0.78 0.81 EU107646 Pygolabis sp. 1 1.00 I49 PT04 Isopoda EU107664 Pygolabis sp. 4 Haloniscus sp. 10 EU364592 0.76 Haloniscus sp. 12 EU364601

0.78 B12 20 1b Isopoda 0.79 IE50 RC08MEJ0076P2T2 1

0.90 EU364622 Haloniscus sp. 21 0.86 G173 Philosciidae S1 100421 G175 Philosciidae S1 LN7657 0.1

34 Molecular systematics of subfauna from the Robe Valley

Figure 10. Neighbour-joining analysis of specimens of Pseudoscrpiones from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

100 IV201 DD14MEB0002-20150607-01sc Pseudoscorpiones Lin 1 98 IV217 RC14MEB0029-20151001-T1-01 Pseudoscorpiones Lin 2 IV206 DD14MRR0008-20150808-T1-02 Pseudoscorpiones Lin 3 58 100 IV209 M2ERC027-t2-03 Pseudoscorpiones Lin 4 IV202 DD14MRR0004T1-04 Pseudoscorpiones Lin 5 IV211 RC13MEA0279-20150807-T1-02 Pseudoscorpiones Lin 6 61 100 IV213 DD14MRR0009-20151002-T2-02 Pseudoscorpiones Lin 7 70 IV219 RC14MEB0101-20151001-T1-02 Pseudoscorpiones Lin 8

100 IV212 DD14MEL0001-20151001-T2-02 Pseudoscorpiones Lin 9 IV226 RC14MRR0020-20151002-T2-01 Pseudoscorpiones Lin 10

100 IV203 DD14MRR0004T1-04 Pseudoscorpiones 100 Lin 11 IV204 DD14MRR0005-T1-05 Pseudoscorpiones 98 IV210 MEARC3814-20150606-02sc Pseudoscorpiones Lin 12 100 IV215 MEARC3814-20150930-T1-01 Pseudoscorpiones IV205 DD14MRR0005-T2-01 Pseudoscorpiones Lin 13 IV208 M2ERC027-t2-03 Pseudoscorpiones Lin 14 100 IV207 DD14MRR0008-20150808-T2-03 Pseudoscorpiones 100 Lin 15 100 IV214 DD14MRR0009-20151002-T3-01 Pseudoscorpiones

0.02

35 Molecular systematics of subfauna from the Robe Valley

Figure 11. Bayesian analysis of COXI haplotypes of Chthoniidae (Pseudoscorpiones) from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

AY156582 Pandinus imperator DQ513111 Siro rubens IV212 DD14MEL0001 20151001 T2 02 lin 9 1.00 IV226 RC14MRR0020 20151002 T2 01 lin 10 1.00 EAP0178 Ophthalmia IS01 MSRC036-20150709-T2-01 0.61 1.00 1.00 IS12 CWRC937-20150912-T1-03 0.86 1.00 IS08 TSOOPP03.20150709-T1-04 Red Hill IS05 KBRC1475-20150708-T2-04 1.00 IS06 CWRC484-20150709-T1-04 EU559513 Austrochtonia sp 0.82 IV203 DD14MRR0004T1 04 lin 11 0.98 1.00 IV210 MEARC3814 20150606 02sc lin 12 IS11 KBRC1533-T1-01 Red Hill 0.66 EU559503 Lagynochthonius johni 0.51 0.97 IS10 KBRCopp15-20150913-02 0.98 G118 Tyrannochthonius

1.00 JIN0078R Lagynochthonius EJR0202 2 9 9 Tyrannochthonius G219 Pseudoscorpian 108935 1.00 0.98 G220 Pseudoscorpian 109177 EU559506 Tyrannochthonius sp. JM 2008

1.00 LB084 Tyrannochthonius 0.53 EXR1148 J20 6 Lagynochthonius 1.00 PI012 Tyrannochthonius 0.98 CM18 RC11KOOD0193P2T1 2 Lagynochthonius sp. 0.94 0.51 GF0173R Lagynochthonius 1.00 G328 LN9199 Tyrannochthonius sp OP G324 101086 Tyrannochthonius sp MA 0.98 EA0270R Lagynochthonius G319 LN8720 Chthoniidae sp MJ 0.1

36 Molecular systematics of subfauna from the Robe Valley

Figure 12. Bayesian analysis of COXI haplotypes of Hyiidae (Pseudoscorpiones) from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

AY156582 Pandinus imperator

DQ513111 Siro rubens

IV201 DD14MEB0002 20150607 01sc lin 1 1.00 IV217 RC14MEB0029 20151001 T1 01 lin 2 0.62 GB0018R Hyiidae Jirrpalpur 1.00 GC0103R Hyiidae Jirrpalpur 0.58 1.00 PSF0090R Indohya Packsaddle

FZ1 Pseudoscorpion RC13KOOD0302P7T1 1

0.88 IV206 DD14MRR0008 20150808 T1 02 lin 3 1.00 1.00 IV209 M2ERC027 t2 03 lin 4 Hyiidae 0.99 G107 Indohya Central Pilbara

0.72 DF18 8668 Indohya sp. Hardey River EU559564 Indohya sp. JM 2008

IV202 DD14MRR0004T1 04 lin 5

0.77 IV211 RC13MEA0279 20150807 T1 02 lin 6 IV213 DD14MRR0009 20151002 T2 02 lin 7 0.63 1.00 IV219 RC14MEB0101 20151001 T1 02 lin 8

IT72 BHRC0512 151013 T3 01 Buckland Hills IV205 DD14MRR0005 T2 01 lin 13 1.00 IV208 M2ERC027 t2 03 lin 14 1.00 1.00 Atemnidae IV207 DD14MRR0008 20150808 T2 03 lin 15

JN018204 Paratemnoides sumatranus Atemnidae

0.1

37 Table 4. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV55 IV81 IV07 IV09 IV14 IV17 IV27 IV61 IV76 IV79 Dbram Dmes Pan Pbyt Pgno Pkry Psp C119 C37 C58 FT4 FT5 FT6 FZ21 GG9 GH7 GH8 HE17 HE4 IS15 IS33 IT67 C17 U72_Shizomid_T95463 U74_Shizomid_T95462 U83_Shizomid_T95450 IV55_RC13MRR0077-T1-02_Schizomida 0.015 0.014 0.013 0.012 0.014 0.012 0.016 0.014 0.014 0.013 0.013 0.015 0.017 0.013 0.015 0.015 0.016 0.014 0.014 0.014 0.013 0.013 0.016 0.014 0.016 0.016 0.015 0.015 0.014 0.015 0.015 0.013 0.013 0.013 0.015 IV81_RC14MRR0017-20151002-T2-01_Schizomida 0.172 0.014 0.014 0.014 0.016 0.014 0.014 0.014 0.013 0.013 0.014 0.014 0.015 0.015 0.014 0.015 0.014 0.014 0.014 0.013 0.013 0.013 0.013 0.014 0.015 0.014 0.015 0.012 0.014 0.013 0.013 0.016 0.014 0.014 0.014 IV07_Budgie20150604-06_Schizomida 0.152 0.153 0.011 0.016 0.014 0.015 0.011 0.012 0.013 0.014 0.014 0.011 0.011 0.011 0.013 0.010 0.014 0.013 0.013 0.014 0.014 0.015 0.014 0.014 0.012 0.012 0.012 0.014 0.012 0.014 0.013 0.013 0.014 0.013 0.012 IV09_DD14MEL0001-t2-01_Schizomida 0.131 0.167 0.108 0.015 0.015 0.014 0.012 0.006 0.014 0.014 0.014 0.012 0.013 0.011 0.012 0.010 0.014 0.012 0.013 0.013 0.012 0.013 0.013 0.012 0.013 0.013 0.012 0.014 0.011 0.013 0.011 0.014 0.014 0.014 0.013 IV14_DD14MRR0005-T1-01_Schizomida 0.123 0.172 0.166 0.153 0.011 0.009 0.015 0.015 0.014 0.016 0.009 0.015 0.016 0.015 0.016 0.017 0.015 0.015 0.013 0.014 0.014 0.014 0.016 0.014 0.015 0.015 0.016 0.013 0.013 0.014 0.015 0.012 0.015 0.014 0.014 IV17_DD14MRR007-20150808-T1-02_Schizomida 0.122 0.150 0.156 0.158 0.091 0.011 0.016 0.014 0.013 0.014 0.011 0.014 0.016 0.014 0.016 0.016 0.014 0.015 0.012 0.013 0.013 0.014 0.015 0.013 0.015 0.014 0.014 0.014 0.013 0.014 0.013 0.014 0.013 0.013 0.014 IV27_M2ERC0053-T2-02_Schizomida 0.119 0.161 0.152 0.147 0.072 0.086 0.015 0.013 0.013 0.013 0.004 0.014 0.015 0.014 0.016 0.016 0.014 0.014 0.013 0.013 0.014 0.013 0.014 0.013 0.015 0.014 0.014 0.014 0.012 0.013 0.015 0.015 0.013 0.013 0.012 IV61_DD14MEC0002-20150929-T3-03_Schizomida 0.163 0.156 0.069 0.117 0.172 0.166 0.167 0.012 0.015 0.015 0.015 0.013 0.002 0.013 0.013 0.011 0.014 0.014 0.011 0.013 0.015 0.014 0.014 0.013 0.014 0.013 0.012 0.014 0.012 0.012 0.012 0.013 0.015 0.014 0.012 IV76_MELUNK11-20151001-T2-01_Schizomida 0.128 0.160 0.111 0.034 0.146 0.152 0.139 0.102 0.013 0.014 0.013 0.012 0.012 0.011 0.011 0.011 0.014 0.012 0.013 0.013 0.012 0.013 0.013 0.012 0.013 0.013 0.013 0.013 0.011 0.013 0.010 0.014 0.014 0.013 0.012 IV79_RC14MEB0060-20151001-T2-01_Schizomida 0.183 0.181 0.156 0.166 0.178 0.172 0.159 0.161 0.160 0.015 0.014 0.015 0.016 0.015 0.015 0.015 0.014 0.015 0.016 0.015 0.014 0.014 0.012 0.014 0.014 0.015 0.013 0.013 0.013 0.014 0.013 0.014 0.014 0.014 0.013 EU272684_Draculoides_bramstokeri 0.152 0.165 0.148 0.135 0.148 0.146 0.144 0.152 0.132 0.155 0.013 0.014 0.015 0.013 0.014 0.012 0.015 0.014 0.012 0.013 0.012 0.013 0.012 0.013 0.014 0.014 0.013 0.014 0.012 0.013 0.012 0.015 0.012 0.013 0.012 EU272730_Draculoides_mesozeirus_SzJ 0.120 0.162 0.149 0.141 0.073 0.089 0.006 0.165 0.136 0.157 0.141 0.014 0.015 0.013 0.016 0.015 0.014 0.015 0.013 0.013 0.014 0.013 0.015 0.014 0.015 0.015 0.015 0.013 0.013 0.013 0.015 0.015 0.013 0.013 0.011 Panachoretus_WAM_T66236_Paradraculoides 0.166 0.160 0.085 0.121 0.165 0.155 0.152 0.113 0.124 0.169 0.147 0.151 0.013 0.012 0.013 0.011 0.015 0.015 0.012 0.014 0.015 0.015 0.017 0.015 0.016 0.015 0.012 0.014 0.014 0.014 0.012 0.014 0.013 0.014 0.012 Pbythius_WAM_T63344_Paradraculoides_byth 0.166 0.161 0.069 0.119 0.171 0.169 0.168 0.002 0.103 0.165 0.156 0.164 0.111 0.013 0.013 0.011 0.014 0.014 0.012 0.014 0.016 0.015 0.014 0.014 0.014 0.014 0.013 0.014 0.012 0.012 0.012 0.014 0.015 0.014 0.013 Pgnophicola_WAM_T63371_3_Paradraculoides 0.131 0.150 0.087 0.095 0.161 0.135 0.148 0.100 0.090 0.174 0.137 0.144 0.121 0.102 0.012 0.009 0.014 0.011 0.013 0.015 0.013 0.016 0.015 0.016 0.013 0.013 0.012 0.013 0.011 0.012 0.011 0.014 0.013 0.013 0.014 Pkryptus_WAM_T65802_Paradraculoides_kryp 0.142 0.165 0.102 0.100 0.150 0.152 0.153 0.110 0.095 0.174 0.140 0.152 0.118 0.108 0.092 0.011 0.014 0.014 0.013 0.015 0.014 0.015 0.014 0.014 0.013 0.013 0.013 0.013 0.013 0.013 0.012 0.014 0.015 0.014 0.013 Psp_OFB_2007_Paradraculoides_sp._OFB-200 0.150 0.156 0.074 0.103 0.166 0.153 0.160 0.089 0.097 0.153 0.137 0.156 0.094 0.090 0.084 0.098 0.015 0.012 0.011 0.015 0.014 0.016 0.015 0.015 0.013 0.014 0.011 0.013 0.013 0.014 0.011 0.014 0.013 0.013 0.013 C119_ex_T95543 0.171 0.144 0.130 0.143 0.170 0.167 0.165 0.137 0.130 0.173 0.162 0.162 0.150 0.137 0.134 0.148 0.147 0.016 0.014 0.014 0.014 0.015 0.015 0.015 0.012 0.011 0.015 0.013 0.014 0.013 0.013 0.016 0.013 0.013 0.012 C37_T92541B 0.144 0.153 0.127 0.127 0.164 0.151 0.153 0.144 0.129 0.186 0.155 0.152 0.142 0.140 0.116 0.123 0.119 0.156 0.012 0.014 0.012 0.015 0.016 0.015 0.014 0.014 0.012 0.012 0.014 0.012 0.012 0.016 0.013 0.013 0.013 C58_T98320 0.150 0.161 0.119 0.130 0.167 0.160 0.163 0.105 0.132 0.172 0.148 0.157 0.150 0.108 0.118 0.123 0.121 0.141 0.120 0.013 0.014 0.015 0.015 0.014 0.013 0.013 0.012 0.011 0.012 0.011 0.012 0.014 0.013 0.012 0.011 FT4_Draculoides_SCH012_PSB0166R 0.169 0.133 0.149 0.160 0.171 0.150 0.160 0.169 0.160 0.161 0.143 0.162 0.148 0.174 0.155 0.171 0.150 0.135 0.175 0.166 0.009 0.006 0.011 0.007 0.014 0.013 0.015 0.014 0.012 0.014 0.013 0.016 0.013 0.013 0.012 FT5_Draculoides_sp_PSB0163R 0.160 0.136 0.146 0.149 0.167 0.161 0.161 0.163 0.146 0.166 0.140 0.159 0.160 0.168 0.139 0.152 0.148 0.132 0.173 0.175 0.064 0.009 0.012 0.009 0.012 0.012 0.014 0.013 0.015 0.015 0.012 0.015 0.012 0.012 0.013 FT6_Draculoides_SCH012_GBP553 0.161 0.138 0.146 0.147 0.164 0.161 0.158 0.163 0.146 0.147 0.143 0.159 0.147 0.168 0.155 0.160 0.144 0.132 0.178 0.175 0.030 0.063 0.011 0.006 0.014 0.014 0.016 0.015 0.014 0.015 0.014 0.016 0.014 0.014 0.013 FZ21_Schizomida_RC13KOOD0308P7T1-1 0.185 0.144 0.142 0.158 0.182 0.170 0.152 0.150 0.149 0.144 0.138 0.152 0.160 0.155 0.145 0.152 0.139 0.149 0.170 0.181 0.122 0.117 0.118 0.011 0.014 0.013 0.013 0.014 0.014 0.013 0.015 0.017 0.014 0.015 0.013 GG9_Draculoides_sp_PSA0806R 0.160 0.133 0.147 0.155 0.164 0.144 0.156 0.167 0.149 0.150 0.135 0.159 0.144 0.173 0.152 0.161 0.142 0.137 0.167 0.171 0.031 0.061 0.028 0.117 0.013 0.014 0.015 0.014 0.013 0.014 0.013 0.016 0.013 0.013 0.013 GH7_Draculoides_Dra-PES16433 0.164 0.146 0.131 0.139 0.175 0.172 0.164 0.146 0.139 0.169 0.162 0.160 0.140 0.148 0.152 0.142 0.145 0.135 0.138 0.133 0.153 0.147 0.147 0.156 0.147 0.007 0.014 0.013 0.013 0.014 0.014 0.014 0.015 0.014 0.014 GH8_Draculoides_Dra-PES16447 0.152 0.150 0.121 0.136 0.155 0.163 0.142 0.146 0.130 0.161 0.160 0.139 0.132 0.147 0.139 0.132 0.145 0.119 0.141 0.130 0.146 0.133 0.136 0.147 0.146 0.038 0.015 0.013 0.013 0.014 0.013 0.014 0.014 0.014 0.014 HE17_BHRC151-20141216-T3-01c 0.151 0.148 0.129 0.124 0.157 0.143 0.148 0.126 0.127 0.175 0.149 0.146 0.141 0.128 0.123 0.111 0.121 0.148 0.115 0.118 0.164 0.156 0.156 0.156 0.159 0.137 0.137 0.012 0.014 0.010 0.012 0.014 0.013 0.013 0.013 HE4_BHRC001-20141028-SC01 0.153 0.146 0.138 0.143 0.149 0.142 0.138 0.137 0.134 0.165 0.165 0.130 0.147 0.137 0.131 0.132 0.132 0.144 0.121 0.118 0.165 0.143 0.160 0.162 0.154 0.145 0.145 0.115 0.013 0.011 0.012 0.014 0.014 0.014 0.013 IS15_KBRC1389_20150909_04_Schizomida 0.161 0.149 0.122 0.127 0.161 0.150 0.135 0.124 0.122 0.155 0.133 0.136 0.142 0.127 0.121 0.127 0.126 0.138 0.135 0.109 0.147 0.166 0.150 0.156 0.144 0.152 0.144 0.124 0.124 0.011 0.012 0.014 0.013 0.014 0.013 IS33_Kbrc116720150809t2_05_Schizomida_sp 0.152 0.142 0.124 0.130 0.155 0.139 0.139 0.121 0.121 0.156 0.130 0.139 0.129 0.118 0.111 0.111 0.121 0.124 0.114 0.108 0.146 0.153 0.149 0.159 0.144 0.138 0.122 0.104 0.099 0.100 0.011 0.015 0.013 0.013 0.013 IT67_BHRC401-151013-T1-01_Schizomida 0.149 0.147 0.114 0.121 0.157 0.146 0.144 0.116 0.111 0.157 0.135 0.141 0.121 0.119 0.116 0.108 0.111 0.130 0.132 0.113 0.155 0.138 0.154 0.144 0.150 0.139 0.127 0.115 0.110 0.099 0.105 0.013 0.013 0.013 0.010 T93233_C17_CBRC099P5T1-3_Paradraculoides_sp_cardo 0.143 0.170 0.117 0.126 0.144 0.130 0.152 0.123 0.134 0.148 0.146 0.150 0.119 0.123 0.115 0.112 0.114 0.161 0.130 0.106 0.166 0.168 0.155 0.162 0.162 0.139 0.125 0.121 0.135 0.116 0.114 0.105 0.014 0.014 0.015 U72_Shizomid_T95463 0.133 0.147 0.124 0.131 0.146 0.121 0.122 0.146 0.132 0.161 0.124 0.120 0.139 0.152 0.129 0.152 0.131 0.129 0.142 0.138 0.130 0.133 0.133 0.147 0.124 0.155 0.149 0.138 0.140 0.133 0.130 0.129 0.134 0.006 0.010 U74_Shizomid_T95462 0.139 0.158 0.127 0.130 0.152 0.133 0.124 0.135 0.129 0.163 0.133 0.122 0.147 0.140 0.135 0.152 0.132 0.129 0.145 0.130 0.139 0.140 0.140 0.150 0.130 0.152 0.150 0.146 0.142 0.130 0.132 0.125 0.137 0.018 0.010 U83_Shizomid_T95450 0.147 0.144 0.124 0.133 0.153 0.144 0.131 0.149 0.130 0.160 0.137 0.125 0.131 0.150 0.140 0.152 0.132 0.137 0.153 0.138 0.127 0.141 0.137 0.142 0.122 0.147 0.146 0.151 0.154 0.145 0.141 0.124 0.141 0.078 0.081 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 9. Estimates of Evolutionary Divergence between Sequences

Lineage IV126 IV92 IV111 IV128 IV108 IV134 IV129 IV100 Nfon Ngle IT06 IT22 IS37 IS38 IS43 IS59 GY19 DH1 L8 107b Nman Ndou B3 60a L26 BF14 sge1m4 2 pm01a cob12 pt03 rr3a nr5miwell IV126 Camp-20150930-02 lin 1 0.011 0.011 0.013 0.013 0.014 0.018 0.019 0.020 0.021 0.008 0.010 0.014 0.013 0.014 0.012 0.011 0.016 0.013 0.016 0.017 0.015 0.017 0.015 0.013 0.016 0.016 0.016 0.008 0.014 IV92 Budgie.20150604-02 lin 2 0.074 0.010 0.014 0.013 0.014 0.017 0.019 0.020 0.020 0.010 0.013 0.014 0.014 0.013 0.009 0.011 0.016 0.013 0.015 0.017 0.014 0.017 0.014 0.014 0.015 0.014 0.016 0.012 0.002 IV111 MB13WARR010.20150606-01 lin 3 0.088 0.077 0.012 0.014 0.014 0.017 0.018 0.019 0.020 0.012 0.013 0.015 0.014 0.014 0.011 0.010 0.016 0.012 0.015 0.015 0.014 0.017 0.015 0.014 0.015 0.015 0.017 0.013 0.013 IV128 DAVES-20150930-04 lin 4 0.101 0.110 0.109 0.014 0.015 0.019 0.020 0.020 0.021 0.011 0.015 0.014 0.015 0.014 0.012 0.013 0.016 0.015 0.017 0.017 0.016 0.018 0.016 0.013 0.016 0.017 0.017 0.014 0.016 IV108 MB13WARR001-20150605-01 lin 5 0.139 0.137 0.147 0.142 0.015 0.018 0.019 0.021 0.021 0.013 0.017 0.013 0.015 0.012 0.014 0.016 0.017 0.016 0.014 0.011 0.011 0.018 0.013 0.016 0.017 0.014 0.013 0.015 0.015 IV134 TOBRC0099-20150930-01 lin 6 0.148 0.147 0.151 0.150 0.158 0.016 0.017 0.018 0.020 0.014 0.015 0.012 0.013 0.016 0.015 0.015 0.017 0.016 0.017 0.017 0.017 0.017 0.015 0.016 0.014 0.015 0.017 0.016 0.016 IV129 DAVES-20150930-04 lin 7 0.302 0.292 0.291 0.303 0.295 0.273 0.013 0.015 0.015 0.018 0.022 0.015 0.016 0.017 0.018 0.018 0.018 0.018 0.021 0.022 0.019 0.019 0.017 0.022 0.020 0.021 0.021 0.020 0.020 IV100 Daves-20150604-01 lin 8 0.322 0.314 0.312 0.341 0.308 0.309 0.155 0.015 0.017 0.020 0.024 0.017 0.018 0.019 0.019 0.019 0.018 0.020 0.020 0.022 0.019 0.018 0.018 0.021 0.020 0.021 0.022 0.021 0.022 KC315635 Niphargus fontanus 0.319 0.314 0.309 0.326 0.322 0.308 0.190 0.205 0.014 0.019 0.026 0.019 0.018 0.020 0.019 0.019 0.019 0.021 0.020 0.024 0.020 0.019 0.018 0.023 0.022 0.023 0.023 0.023 0.023 KC315646 Niphargus glenniei 0.330 0.322 0.315 0.330 0.326 0.328 0.190 0.213 0.167 0.020 0.026 0.020 0.020 0.021 0.022 0.021 0.020 0.021 0.019 0.022 0.019 0.020 0.020 0.022 0.021 0.023 0.022 0.023 0.022 IT06 BHRC118.20151013-02 0.034 0.060 0.077 0.097 0.135 0.148 0.306 0.330 0.324 0.333 0.010 0.013 0.012 0.013 0.011 0.011 0.016 0.013 0.015 0.015 0.013 0.016 0.015 0.012 0.015 0.016 0.015 0.010 0.012 IT22 BHRC231.20151014-03 0.047 0.074 0.079 0.108 0.140 0.145 0.322 0.348 0.348 0.338 0.045 0.017 0.018 0.016 0.015 0.014 0.021 0.014 0.018 0.017 0.016 0.021 0.018 0.016 0.017 0.017 0.017 0.011 0.014 IS37 RHWB005 20150910 01 0.155 0.155 0.155 0.153 0.161 0.103 0.273 0.306 0.326 0.326 0.151 0.164 0.014 0.013 0.014 0.014 0.016 0.015 0.018 0.016 0.015 0.017 0.016 0.017 0.013 0.016 0.017 0.016 0.016 IS38 RHWB005 20150910 01 0.166 0.151 0.162 0.166 0.172 0.114 0.269 0.311 0.326 0.326 0.159 0.177 0.110 0.016 0.013 0.014 0.015 0.015 0.017 0.017 0.015 0.015 0.015 0.016 0.013 0.018 0.019 0.015 0.015 IS43 RHWB006 20150910 01 0.144 0.136 0.144 0.147 0.099 0.186 0.297 0.311 0.325 0.331 0.134 0.129 0.158 0.188 0.014 0.014 0.017 0.015 0.017 0.013 0.011 0.020 0.015 0.015 0.018 0.016 0.015 0.015 0.016 IS59 KBRC1310 20150911 02 0.104 0.084 0.093 0.107 0.153 0.175 0.292 0.322 0.326 0.330 0.101 0.103 0.169 0.169 0.148 0.011 0.014 0.013 0.015 0.017 0.015 0.016 0.015 0.012 0.015 0.016 0.017 0.013 0.012 GY19 BIL4nr-20141109-20 0.078 0.077 0.081 0.110 0.145 0.160 0.270 0.305 0.318 0.316 0.072 0.080 0.149 0.145 0.134 0.101 0.015 0.005 0.016 0.017 0.015 0.016 0.015 0.013 0.017 0.017 0.017 0.013 0.014 DH1 BIL4nr-1112-03-01 0.166 0.169 0.167 0.155 0.188 0.151 0.291 0.309 0.317 0.319 0.163 0.179 0.137 0.151 0.196 0.175 0.155 0.017 0.016 0.018 0.017 0.007 0.016 0.018 0.017 0.018 0.019 0.018 0.019 L8 107b Nedsia sp 2 0.082 0.086 0.086 0.124 0.143 0.163 0.283 0.323 0.324 0.317 0.077 0.076 0.151 0.152 0.138 0.111 0.016 0.170 0.016 0.017 0.015 0.017 0.016 0.014 0.017 0.017 0.017 0.013 0.015 Norcapensis mandibulis 0.168 0.159 0.152 0.176 0.182 0.168 0.294 0.292 0.315 0.294 0.165 0.168 0.171 0.168 0.197 0.173 0.165 0.170 0.163 0.016 0.016 0.017 0.016 0.018 0.018 0.018 0.017 0.016 0.017 wc17 1 Nedsia douglasi 0.149 0.149 0.142 0.140 0.068 0.153 0.297 0.310 0.323 0.312 0.136 0.142 0.161 0.178 0.096 0.159 0.144 0.185 0.142 0.166 0.008 0.019 0.015 0.015 0.017 0.014 0.012 0.017 0.017 B3 60a Nedsia sp 1 0.152 0.154 0.151 0.145 0.081 0.156 0.320 0.330 0.337 0.337 0.139 0.152 0.170 0.173 0.114 0.164 0.158 0.197 0.154 0.180 0.038 0.018 0.014 0.015 0.017 0.013 0.012 0.016 0.016 L26 S5-200 unknown sp 3 0.175 0.171 0.175 0.168 0.189 0.152 0.289 0.311 0.315 0.317 0.165 0.182 0.151 0.152 0.202 0.178 0.159 0.029 0.165 0.168 0.178 0.187 0.016 0.018 0.016 0.019 0.019 0.018 0.019 BF14 X62P2-2011-01 0.183 0.180 0.181 0.186 0.161 0.156 0.277 0.294 0.324 0.313 0.187 0.195 0.154 0.149 0.178 0.188 0.173 0.169 0.186 0.176 0.174 0.175 0.171 0.017 0.015 0.016 0.019 0.017 0.016 sge1m4 2 0.114 0.101 0.099 0.114 0.161 0.165 0.292 0.338 0.334 0.326 0.095 0.117 0.169 0.163 0.148 0.089 0.082 0.186 0.082 0.171 0.147 0.152 0.178 0.199 0.016 0.018 0.017 0.013 0.014 pm01a 10 0.153 0.137 0.149 0.141 0.164 0.105 0.275 0.300 0.303 0.317 0.137 0.148 0.099 0.120 0.162 0.160 0.155 0.147 0.158 0.164 0.157 0.168 0.139 0.166 0.159 0.017 0.017 0.016 0.015 cob12 5a 0.149 0.134 0.126 0.155 0.105 0.155 0.284 0.313 0.330 0.309 0.141 0.136 0.153 0.181 0.128 0.153 0.145 0.187 0.134 0.162 0.115 0.128 0.183 0.174 0.163 0.164 0.011 0.016 0.014 pt03 11a 0.153 0.147 0.143 0.162 0.101 0.170 0.305 0.330 0.334 0.319 0.145 0.151 0.172 0.187 0.128 0.158 0.151 0.195 0.149 0.160 0.106 0.118 0.187 0.181 0.167 0.174 0.067 0.016 0.016 rr3a 2b 0.027 0.076 0.095 0.113 0.139 0.139 0.298 0.328 0.321 0.324 0.040 0.054 0.158 0.164 0.139 0.101 0.080 0.164 0.082 0.172 0.146 0.145 0.164 0.202 0.106 0.149 0.151 0.155 0.012 nr5miwelld 0.086 0.002 0.086 0.120 0.139 0.139 0.290 0.321 0.324 0.315 0.067 0.080 0.158 0.155 0.137 0.092 0.086 0.181 0.090 0.155 0.149 0.147 0.170 0.195 0.099 0.134 0.137 0.149 0.078 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 12. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV139 IV140 IV141 IV143 IS87 Mmult Gbon Gflav Slin Ssex Pmon AK10 BQ12 BX11 BX12 BX13 BX8 BX9 CQ2 CQ3 CQ5 CQ6 EY5 G123 G124 G349 IV139_DD14MRR0004T1-01_Chilopoda 0.000 0.002 0.017 0.019 0.018 0.019 0.017 0.019 0.018 0.019 0.021 0.020 0.018 0.020 0.020 0.020 0.019 0.021 0.017 0.019 0.017 0.020 0.019 0.019 0.019 IV140_DD14MRR0004T1-01_Chilopoda 0.000 0.002 0.017 0.019 0.018 0.019 0.017 0.019 0.018 0.019 0.021 0.020 0.018 0.020 0.020 0.020 0.019 0.021 0.017 0.019 0.017 0.020 0.019 0.019 0.019 IV141_DD14MRR0005-T1-04_Chilopoda 0.003 0.003 0.017 0.018 0.018 0.019 0.017 0.019 0.018 0.019 0.021 0.020 0.017 0.020 0.020 0.020 0.018 0.021 0.017 0.018 0.017 0.019 0.019 0.019 0.019 IV143_MEARC4383-20151001-T2-01_Chilopoda 0.229 0.229 0.228 0.017 0.018 0.020 0.018 0.019 0.017 0.018 0.017 0.018 0.018 0.017 0.019 0.019 0.017 0.017 0.017 0.015 0.017 0.018 0.019 0.016 0.018 IS87_KBRC136220150911T2_03_Geophilida_sp 0.227 0.227 0.225 0.219 0.014 0.018 0.018 0.018 0.016 0.019 0.018 0.017 0.017 0.017 0.019 0.019 0.018 0.016 0.017 0.017 0.018 0.018 0.017 0.017 0.017 AB610774_Mecistocephalus_multidentatus 0.191 0.191 0.190 0.229 0.188 0.020 0.016 0.017 0.017 0.017 0.018 0.018 0.019 0.018 0.021 0.021 0.019 0.017 0.019 0.020 0.017 0.021 0.018 0.018 0.019 KF569297_Gnathoribautia_bonensis 0.206 0.206 0.206 0.235 0.239 0.202 0.020 0.020 0.019 0.018 0.018 0.017 0.019 0.021 0.021 0.021 0.022 0.022 0.021 0.021 0.022 0.022 0.020 0.020 0.019 JN306685_Geophilus_flavus 0.223 0.223 0.220 0.281 0.231 0.217 0.194 0.016 0.018 0.020 0.019 0.019 0.019 0.019 0.020 0.020 0.021 0.018 0.019 0.019 0.019 0.019 0.019 0.018 0.018 KR736251_Stenotaenia_linearis 0.245 0.245 0.243 0.271 0.264 0.240 0.216 0.196 0.021 0.018 0.020 0.019 0.021 0.020 0.020 0.020 0.020 0.018 0.018 0.020 0.018 0.022 0.019 0.018 0.018 AY288745_Scolopocryptops_sexspinosus_Cryptopidae 0.221 0.221 0.220 0.218 0.220 0.223 0.225 0.235 0.244 0.017 0.018 0.018 0.016 0.019 0.020 0.021 0.019 0.018 0.019 0.019 0.017 0.017 0.019 0.019 0.017 AF334321_Paralamyctes_monteithi_Henicopidae 0.191 0.191 0.189 0.210 0.232 0.198 0.205 0.254 0.242 0.206 0.018 0.019 0.019 0.018 0.020 0.019 0.019 0.020 0.019 0.017 0.019 0.020 0.020 0.019 0.018 AK10_PE111022_Cryptop_DC10 0.252 0.252 0.251 0.211 0.239 0.250 0.229 0.245 0.272 0.218 0.213 0.017 0.018 0.018 0.018 0.018 0.019 0.017 0.018 0.017 0.016 0.019 0.019 0.018 0.018 BQ12_PES_5087_Cryptops_won1 0.232 0.232 0.232 0.215 0.225 0.226 0.227 0.237 0.262 0.190 0.213 0.198 0.019 0.019 0.020 0.019 0.019 0.018 0.017 0.020 0.019 0.022 0.020 0.017 0.015 BX11_PES-0968_Cryptops_pilbara1 0.250 0.250 0.246 0.227 0.230 0.240 0.241 0.243 0.268 0.214 0.224 0.194 0.213 0.017 0.019 0.019 0.018 0.017 0.018 0.018 0.016 0.010 0.019 0.019 0.018 BX12_PES-3783_Cryptops_pilbara2 0.234 0.234 0.233 0.209 0.231 0.229 0.249 0.260 0.260 0.240 0.247 0.219 0.233 0.169 0.017 0.017 0.018 0.016 0.016 0.017 0.015 0.018 0.018 0.017 0.018 BX13_991-5262_Cryptops_MH1 0.237 0.237 0.237 0.230 0.248 0.246 0.260 0.271 0.271 0.239 0.251 0.218 0.236 0.201 0.184 0.003 0.017 0.017 0.015 0.018 0.018 0.023 0.019 0.019 0.019 BX8_PES-5889_Cryptops_MH2 0.239 0.239 0.239 0.225 0.248 0.248 0.262 0.272 0.272 0.244 0.253 0.215 0.233 0.201 0.181 0.006 0.017 0.017 0.014 0.018 0.018 0.023 0.018 0.019 0.019 BX9_PES-5272_Cryptops_MH2 0.232 0.232 0.229 0.204 0.231 0.247 0.255 0.271 0.266 0.213 0.215 0.208 0.218 0.173 0.186 0.175 0.175 0.018 0.019 0.019 0.018 0.018 0.018 0.020 0.019 CQ2_nr_Cryptops_sp._B15_CCWUNK05 0.246 0.246 0.246 0.198 0.225 0.228 0.238 0.240 0.283 0.230 0.249 0.207 0.192 0.210 0.194 0.228 0.222 0.228 0.015 0.018 0.015 0.018 0.017 0.018 0.016 CQ3_Cryptops_sp._B32_CC1798 0.227 0.227 0.227 0.220 0.229 0.229 0.246 0.246 0.254 0.220 0.239 0.211 0.198 0.193 0.174 0.127 0.124 0.171 0.197 0.017 0.016 0.020 0.019 0.019 0.016 CQ5_nr_Cryptops_sp._B11_PCRC088 0.236 0.236 0.233 0.180 0.225 0.236 0.233 0.257 0.272 0.220 0.191 0.204 0.194 0.209 0.218 0.220 0.225 0.229 0.185 0.205 0.016 0.021 0.018 0.017 0.016 CQ6_Cryptops_sp._B30_1106_13_EX13 0.227 0.227 0.228 0.220 0.244 0.246 0.255 0.264 0.271 0.211 0.227 0.193 0.220 0.178 0.153 0.178 0.175 0.167 0.204 0.164 0.205 0.019 0.017 0.018 0.018 EY5_Cryptops_sp_B35 0.241 0.241 0.238 0.202 0.229 0.241 0.249 0.236 0.268 0.217 0.230 0.197 0.225 0.062 0.157 0.213 0.210 0.150 0.208 0.180 0.218 0.176 0.021 0.021 0.019 G123_Cryptops_Upper_South_Fortescue 0.235 0.235 0.232 0.234 0.239 0.246 0.223 0.243 0.267 0.207 0.216 0.222 0.237 0.242 0.236 0.257 0.254 0.250 0.236 0.248 0.230 0.213 0.261 0.019 0.017 G124_Cryptops_Upper_South_Fortescue 0.242 0.242 0.241 0.204 0.236 0.236 0.232 0.265 0.271 0.216 0.214 0.220 0.170 0.227 0.236 0.228 0.231 0.225 0.209 0.220 0.182 0.221 0.225 0.241 0.016 G349_Cryptopidae_sp_MJ_LN9808 0.240 0.240 0.240 0.211 0.219 0.243 0.224 0.245 0.280 0.211 0.210 0.203 0.158 0.220 0.239 0.231 0.228 0.223 0.192 0.195 0.182 0.217 0.227 0.226 0.166 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. Disclaimer:The number Although of base utmost differences care has been per taken site to fromensure between the correctness sequences of the caption, are shown. the caption Standard text is provided error "asestimate(s) is" without any are warranty shown of above any kind. the Authors diagonal. advise Thethe user analysis to carefully involved check the 42 caption nucleotide prior to sequences.its use for any purposeCodon and positions report any included errors or problemswere 1st+2nd+3rd+Noncoding. to the authors immediately All (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 15. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV144 IV145 IV146 IV147 IV148 IV150 IV151 IV152 IV153 FH1 J11-1 BX6 BX7 G344 G348 CF5 CF6 CF7 CF8 CF9 CF10 CF13 CF17 CF18+Y1:Y31edited_JIN0968R_Zuphiini_sp._Carabidae Nvio Pmut Hdis Stob Eibl Tsp Kalm IV144_DD14MRR0004T1-03_Coleoptera 0.000 0.000 0.000 0.016 0.015 0.015 0.016 0.016 0.017 0.018 0.020 0.020 0.019 0.018 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.015 0.014 0.015 0.016 0.017 0.017 0.015 0.016 IV145_DD14MRR0004T1-03_Coleoptera 0.000 0.000 0.000 0.016 0.015 0.015 0.016 0.016 0.017 0.018 0.020 0.020 0.019 0.018 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.015 0.014 0.015 0.016 0.017 0.017 0.015 0.016 IV146_DD14MRR0004T1-03_Coleoptera 0.000 0.000 0.000 0.016 0.015 0.015 0.016 0.016 0.017 0.018 0.020 0.020 0.019 0.018 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.015 0.014 0.015 0.016 0.017 0.017 0.015 0.016 IV147_DD14MRR0004T1-03_Coleoptera 0.000 0.000 0.000 0.016 0.015 0.015 0.016 0.016 0.017 0.018 0.020 0.020 0.019 0.018 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.015 0.014 0.015 0.016 0.017 0.017 0.015 0.016 IV148_DD14MRR009-20150808-T3-01_Coleoptera 0.240 0.240 0.240 0.240 0.016 0.017 0.016 0.015 0.015 0.019 0.021 0.021 0.020 0.019 0.013 0.014 0.014 0.014 0.015 0.015 0.015 0.018 0.015 0.015 0.017 0.016 0.016 0.019 0.017 0.015 IV150_RC14MEB0060-20151001-T2-02_Coleoptera 0.258 0.258 0.258 0.258 0.190 0.015 0.007 0.014 0.016 0.018 0.019 0.020 0.018 0.020 0.015 0.015 0.015 0.015 0.015 0.016 0.016 0.016 0.017 0.016 0.016 0.018 0.015 0.016 0.014 0.015 IV151_MEARC3814-20150807-T1-02_Coleoptera 0.198 0.198 0.198 0.198 0.205 0.234 0.017 0.015 0.018 0.017 0.015 0.016 0.016 0.020 0.015 0.015 0.014 0.014 0.015 0.015 0.015 0.016 0.015 0.016 0.013 0.012 0.016 0.018 0.016 0.014 IV152_MEARC4400-t1-01_Coleoptera 0.271 0.271 0.271 0.271 0.190 0.032 0.234 0.015 0.016 0.019 0.019 0.021 0.019 0.020 0.015 0.014 0.015 0.015 0.015 0.015 0.015 0.017 0.017 0.017 0.017 0.018 0.016 0.017 0.016 0.015 IV153_MEARC4400-t1-01_Coleoptera 0.211 0.211 0.211 0.211 0.215 0.239 0.198 0.236 0.016 0.015 0.020 0.018 0.019 0.018 0.017 0.017 0.017 0.017 0.016 0.017 0.017 0.014 0.012 0.015 0.016 0.015 0.003 0.015 0.015 0.016 FH1_Curculionidae_Genus2_B15_EW0540 0.236 0.236 0.236 0.236 0.202 0.231 0.225 0.234 0.234 0.017 0.019 0.019 0.020 0.020 0.014 0.014 0.014 0.014 0.013 0.013 0.013 0.015 0.014 0.017 0.018 0.018 0.016 0.015 0.014 0.015 PSF0019R_LN6326_J11-1_Zuphiinae_S1_Carabidae 0.216 0.216 0.216 0.216 0.249 0.268 0.197 0.274 0.232 0.211 0.023 0.021 0.021 0.017 0.017 0.017 0.016 0.017 0.016 0.016 0.017 0.004 0.013 0.016 0.017 0.015 0.016 0.018 0.017 0.019 BX6_PES-5290_Bembidion_MH1_CaK_Carabidae 0.222 0.222 0.222 0.222 0.243 0.300 0.197 0.310 0.222 0.262 0.202 0.008 0.018 0.017 0.018 0.019 0.018 0.018 0.018 0.019 0.020 0.019 0.017 0.017 0.016 0.015 0.019 0.019 0.020 0.018 BX7_PES-5285_Bembidion_MH1_CaK_Carabidae 0.225 0.225 0.225 0.225 0.239 0.296 0.186 0.303 0.216 0.255 0.190 0.031 0.017 0.016 0.019 0.020 0.018 0.018 0.019 0.019 0.020 0.018 0.016 0.017 0.017 0.015 0.018 0.020 0.021 0.020 G344_LN9362_Anillini_sp_OP_Carabidae 0.211 0.211 0.211 0.211 0.217 0.274 0.164 0.269 0.211 0.247 0.196 0.160 0.152 0.017 0.018 0.018 0.018 0.019 0.019 0.019 0.019 0.018 0.017 0.017 0.014 0.013 0.019 0.021 0.020 0.019 G348_LN8860_Zuphiini_sp_UM_Carabidae 0.236 0.236 0.236 0.236 0.261 0.287 0.202 0.280 0.216 0.224 0.116 0.198 0.180 0.189 0.019 0.020 0.020 0.020 0.019 0.019 0.020 0.014 0.014 0.018 0.020 0.017 0.018 0.021 0.023 0.022 CF5_GB0002R_Curculionidae_Genus_1_sp._B06 0.220 0.220 0.220 0.220 0.173 0.224 0.204 0.228 0.214 0.199 0.231 0.217 0.219 0.234 0.228 0.007 0.008 0.008 0.006 0.007 0.007 0.015 0.014 0.016 0.015 0.014 0.017 0.017 0.016 0.014 CF6_GF0421R_Curculionidae_Genus_1_sp._B02 0.217 0.217 0.217 0.217 0.180 0.217 0.196 0.218 0.214 0.197 0.230 0.226 0.223 0.242 0.228 0.036 0.007 0.006 0.006 0.006 0.005 0.015 0.015 0.016 0.015 0.015 0.017 0.017 0.016 0.013 CF7_PSC0890R_Curculionidae_Genus_1_sp._B02 0.223 0.223 0.223 0.223 0.177 0.215 0.193 0.223 0.206 0.200 0.233 0.228 0.217 0.230 0.233 0.046 0.045 0.004 0.007 0.008 0.007 0.015 0.015 0.016 0.015 0.014 0.018 0.016 0.015 0.013 CF8_PSD0107R_Curculionidae_Genus_1_sp._B08 0.220 0.220 0.220 0.220 0.177 0.217 0.201 0.224 0.209 0.199 0.236 0.235 0.223 0.234 0.239 0.042 0.038 0.013 0.007 0.007 0.006 0.015 0.015 0.016 0.015 0.014 0.018 0.016 0.016 0.013 CF9_SF0034R_Curculionidae_Genus_1_sp._B07 0.215 0.215 0.215 0.215 0.167 0.209 0.190 0.214 0.208 0.188 0.224 0.223 0.221 0.238 0.226 0.029 0.024 0.041 0.039 0.004 0.004 0.015 0.015 0.016 0.014 0.015 0.017 0.017 0.015 0.013 CF10_SF0139R_Curculionidae_Genus_1_sp._B07 0.214 0.214 0.214 0.214 0.176 0.220 0.192 0.221 0.209 0.188 0.219 0.228 0.221 0.240 0.226 0.038 0.024 0.039 0.035 0.014 0.004 0.015 0.015 0.017 0.015 0.016 0.017 0.018 0.015 0.014 CF13_SF0725R_Curculionidae_Genus_1_sp._B09 0.218 0.218 0.218 0.218 0.171 0.220 0.196 0.218 0.208 0.192 0.224 0.221 0.223 0.238 0.230 0.035 0.021 0.038 0.033 0.014 0.014 0.015 0.015 0.016 0.015 0.015 0.017 0.018 0.016 0.014 CF17_PSC0889R_Zuphiini_sp_B03_Carabidae 0.220 0.220 0.220 0.220 0.231 0.256 0.183 0.258 0.220 0.196 0.010 0.179 0.170 0.175 0.115 0.216 0.214 0.221 0.223 0.207 0.206 0.206 0.012 0.015 0.016 0.014 0.015 0.018 0.017 0.018 CF18_edited_JIN0968R_Zuphiini_sp._Carabidae 0.220 0.220 0.220 0.220 0.211 0.249 0.184 0.249 0.209 0.188 0.124 0.195 0.173 0.188 0.117 0.196 0.199 0.195 0.197 0.188 0.192 0.193 0.114 0.015 0.016 0.013 0.013 0.016 0.017 0.015 KJ961916_Necrobia_violacea_voucher_ZMUO_Cleridae 0.166 0.166 0.166 0.166 0.202 0.240 0.175 0.243 0.170 0.202 0.205 0.213 0.203 0.176 0.222 0.210 0.205 0.208 0.208 0.201 0.202 0.201 0.202 0.193 0.015 0.013 0.015 0.016 0.016 0.015 EU710801_Pterostichus_mutus_Carabidae 0.189 0.189 0.189 0.189 0.204 0.247 0.124 0.251 0.210 0.227 0.188 0.184 0.188 0.150 0.226 0.194 0.191 0.207 0.210 0.182 0.189 0.188 0.183 0.213 0.166 0.012 0.016 0.020 0.015 0.016 HM180603_Harpalus_discrepans_Carabidae 0.182 0.182 0.182 0.182 0.170 0.237 0.135 0.237 0.174 0.221 0.185 0.174 0.174 0.128 0.204 0.176 0.180 0.177 0.179 0.170 0.177 0.173 0.169 0.180 0.150 0.106 0.015 0.018 0.017 0.015 KJ962132_Stricticollis_tobias_Anthicidae 0.219 0.219 0.219 0.219 0.223 0.249 0.205 0.246 0.009 0.243 0.238 0.225 0.222 0.210 0.227 0.220 0.220 0.210 0.213 0.214 0.216 0.214 0.231 0.214 0.166 0.216 0.181 0.015 0.015 0.016 GU213687_Echinodera_ibleiensis_Curculionidae 0.252 0.252 0.252 0.252 0.214 0.190 0.223 0.195 0.222 0.219 0.272 0.284 0.284 0.256 0.255 0.208 0.202 0.198 0.199 0.205 0.208 0.207 0.266 0.225 0.216 0.245 0.222 0.223 0.016 0.016 HE615891_Trigonopterus_sp._Curculionidae 0.250 0.250 0.250 0.250 0.224 0.196 0.215 0.204 0.216 0.205 0.246 0.298 0.284 0.263 0.267 0.209 0.202 0.190 0.201 0.192 0.193 0.195 0.244 0.231 0.218 0.224 0.218 0.218 0.198 0.016 EU286486_Kyklioacalles_almadensis_Curculionidae 0.237 0.237 0.237 0.237 0.159 0.226 0.181 0.228 0.215 0.181 0.214 0.231 0.217 0.212 0.212 0.175 0.182 0.182 0.185 0.171 0.182 0.179 0.203 0.182 0.182 0.191 0.191 0.219 0.193 0.212 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 19. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV154_DD11MEB001T1-01_Diplura IV155_DD11MEB001T1-01_Diplura IV156_DD11MEC0005-02_Diplura IV157_DD14MEB0005-20151001-T3-02_Diplura IV159_RC14MEB0101-20151001-T2-01_Diplura Dsp Jsol Ctil Lweb BX29 BX30 BX40 BX16 BX17 G473 BQ7 BQ10 G333 G334 G336 BQ11 BQ8 G432 G433 G329 G330 G331 G332 IT74 IV154_DD11MEB001T1-01_Diplura 0.014 0.016 0.013 0.013 0.015 0.015 0.015 0.017 0.016 0.016 0.016 0.014 0.015 0.016 0.017 0.017 0.018 0.017 0.017 0.017 0.015 0.015 0.017 0.015 0.015 0.015 0.016 0.016 IV155_DD11MEB001T1-01_Diplura 0.144 0.015 0.009 0.008 0.015 0.016 0.016 0.017 0.016 0.017 0.016 0.013 0.015 0.015 0.017 0.017 0.018 0.017 0.017 0.018 0.014 0.016 0.015 0.015 0.016 0.014 0.016 0.014 IV156_DD11MEC0005-02_Diplura 0.261 0.256 0.014 0.015 0.016 0.013 0.018 0.016 0.012 0.014 0.016 0.013 0.016 0.017 0.015 0.015 0.018 0.018 0.015 0.017 0.015 0.013 0.015 0.015 0.015 0.013 0.013 0.014 IV157_DD14MEB0005-20151001-T3-02_Diplura 0.153 0.054 0.243 0.003 0.014 0.015 0.016 0.017 0.016 0.017 0.015 0.013 0.014 0.015 0.016 0.016 0.017 0.016 0.015 0.016 0.014 0.017 0.016 0.016 0.016 0.014 0.016 0.014 IV159_RC14MEB0101-20151001-T2-01_Diplura 0.153 0.047 0.241 0.006 0.014 0.015 0.016 0.017 0.016 0.017 0.015 0.013 0.014 0.015 0.016 0.016 0.017 0.016 0.015 0.016 0.014 0.017 0.015 0.016 0.016 0.014 0.016 0.014 Diplura_sp._HQ943342_DA 0.249 0.227 0.263 0.223 0.222 0.014 0.014 0.015 0.018 0.017 0.015 0.015 0.015 0.015 0.016 0.015 0.017 0.017 0.016 0.016 0.015 0.016 0.016 0.015 0.015 0.017 0.016 0.014 Japyx_solifugus_AY771989_DC 0.257 0.254 0.207 0.239 0.242 0.243 0.017 0.017 0.013 0.013 0.016 0.015 0.016 0.017 0.017 0.019 0.016 0.017 0.016 0.018 0.014 0.015 0.015 0.013 0.013 0.014 0.014 0.016 AF370844_Campodea_tillyardi 0.219 0.222 0.281 0.216 0.214 0.214 0.260 0.017 0.016 0.016 0.018 0.017 0.017 0.017 0.017 0.017 0.017 0.018 0.019 0.018 0.018 0.015 0.017 0.016 0.017 0.019 0.017 0.018 HQ882832_Lepidocampa_weberi 0.253 0.211 0.263 0.213 0.215 0.233 0.250 0.233 0.016 0.017 0.016 0.014 0.015 0.015 0.016 0.015 0.019 0.017 0.014 0.016 0.017 0.018 0.016 0.016 0.018 0.016 0.017 0.018 BX29_PES-5141_Parajapygidae_MH1_DS 0.273 0.270 0.175 0.258 0.258 0.263 0.222 0.278 0.247 0.007 0.016 0.016 0.014 0.016 0.016 0.013 0.018 0.017 0.015 0.017 0.014 0.013 0.014 0.015 0.014 0.012 0.015 0.016 BX30_PES-5190_Parajapygidae_MH1_DS 0.276 0.268 0.178 0.255 0.255 0.267 0.226 0.274 0.244 0.030 0.016 0.016 0.015 0.017 0.017 0.015 0.019 0.017 0.016 0.018 0.015 0.013 0.014 0.014 0.013 0.011 0.014 0.017 BX40_PES-5878_Projapygidae_MH1_DT 0.283 0.295 0.274 0.281 0.278 0.275 0.257 0.287 0.289 0.270 0.280 0.016 0.016 0.016 0.012 0.013 0.014 0.014 0.012 0.013 0.015 0.016 0.016 0.015 0.015 0.016 0.017 0.018 BX16_PES-5195_Japygidae_MH1_DQ 0.229 0.215 0.203 0.210 0.210 0.220 0.179 0.249 0.204 0.213 0.219 0.235 0.011 0.016 0.013 0.015 0.018 0.017 0.015 0.016 0.011 0.013 0.013 0.013 0.015 0.014 0.015 0.014 BX17_PES-5952_Japygidae_MH1_DR 0.252 0.237 0.204 0.223 0.224 0.248 0.200 0.267 0.213 0.204 0.208 0.262 0.130 0.016 0.014 0.015 0.017 0.017 0.015 0.016 0.013 0.015 0.013 0.014 0.014 0.015 0.016 0.013 G473_111138b_Anajapygidae_sp_NS_DB 0.226 0.251 0.242 0.248 0.247 0.245 0.236 0.252 0.257 0.244 0.253 0.253 0.215 0.232 0.016 0.017 0.018 0.017 0.017 0.018 0.018 0.016 0.017 0.018 0.014 0.017 0.016 0.017 BQ7_PES_4269_Projapygidae_DK 0.285 0.267 0.246 0.253 0.251 0.287 0.278 0.282 0.285 0.251 0.256 0.168 0.235 0.234 0.258 0.010 0.014 0.014 0.012 0.013 0.016 0.016 0.016 0.017 0.016 0.017 0.016 0.017 BQ10_PES_2546_Projapygidae_DL 0.292 0.285 0.245 0.262 0.264 0.283 0.265 0.280 0.286 0.246 0.251 0.168 0.246 0.248 0.258 0.113 0.014 0.015 0.011 0.014 0.015 0.015 0.015 0.016 0.017 0.015 0.016 0.017 G333_LN8949_Projapygidae_sp_MJ_DN 0.290 0.298 0.270 0.281 0.284 0.278 0.251 0.283 0.283 0.265 0.264 0.187 0.248 0.241 0.267 0.170 0.187 0.013 0.013 0.014 0.018 0.017 0.019 0.016 0.016 0.017 0.018 0.017 G334_LN9141_Projapygidae_sp_UM_DO 0.287 0.297 0.262 0.283 0.280 0.269 0.254 0.275 0.269 0.270 0.274 0.168 0.246 0.258 0.276 0.183 0.181 0.141 0.012 0.014 0.016 0.017 0.018 0.017 0.017 0.017 0.017 0.017 G336_LN9557_Projapygidae_sp_OP_DM 0.271 0.289 0.251 0.265 0.265 0.254 0.246 0.278 0.277 0.242 0.254 0.168 0.236 0.243 0.255 0.143 0.134 0.160 0.145 0.012 0.014 0.016 0.015 0.014 0.015 0.015 0.016 0.017 BQ11_PES_5439_Projapygidae_won1_DP 0.271 0.300 0.249 0.286 0.281 0.255 0.233 0.246 0.268 0.257 0.270 0.143 0.220 0.238 0.242 0.156 0.162 0.156 0.152 0.146 0.016 0.016 0.016 0.017 0.016 0.018 0.017 0.018 BQ8_PES_4283_Japygidae_won1_DE 0.248 0.233 0.185 0.226 0.224 0.216 0.185 0.251 0.230 0.210 0.217 0.252 0.138 0.147 0.215 0.247 0.252 0.255 0.242 0.242 0.233 0.015 0.013 0.011 0.015 0.015 0.016 0.014 G432_110953_Japygidae_sp_Central_Pilbara_DH 0.255 0.263 0.205 0.246 0.245 0.247 0.202 0.270 0.252 0.199 0.217 0.272 0.157 0.149 0.232 0.252 0.260 0.280 0.258 0.255 0.245 0.172 0.013 0.014 0.014 0.016 0.015 0.013 G433_110848_Japygidae_Central_Pilbara_sp_DG 0.261 0.247 0.191 0.227 0.226 0.244 0.190 0.259 0.227 0.204 0.213 0.247 0.128 0.153 0.226 0.237 0.247 0.258 0.256 0.228 0.231 0.142 0.169 0.014 0.015 0.014 0.015 0.014 G329_LN9310_Japygidae_sp_MA_DF 0.255 0.251 0.194 0.241 0.238 0.237 0.186 0.245 0.233 0.215 0.211 0.234 0.137 0.163 0.228 0.256 0.255 0.255 0.251 0.248 0.249 0.112 0.183 0.158 0.011 0.015 0.015 0.015 G330_LN8732_Japygidae_sp_MJ_DD 0.247 0.234 0.218 0.229 0.227 0.230 0.168 0.253 0.251 0.204 0.193 0.241 0.171 0.185 0.205 0.246 0.237 0.241 0.267 0.226 0.227 0.181 0.182 0.163 0.161 0.013 0.014 0.015 G331_LN9880_Parajapygidae_sp_MJ_DI 0.286 0.279 0.190 0.268 0.271 0.267 0.217 0.293 0.282 0.152 0.151 0.272 0.210 0.219 0.261 0.275 0.267 0.252 0.274 0.259 0.271 0.204 0.231 0.204 0.219 0.223 0.012 0.017 G332_LN8894_Parajapygidae_sp_MJ_DJ 0.277 0.265 0.187 0.255 0.257 0.257 0.219 0.290 0.263 0.174 0.164 0.283 0.203 0.213 0.245 0.273 0.267 0.252 0.262 0.255 0.273 0.211 0.220 0.201 0.205 0.209 0.152 0.016 IT74_BHRC0514-151013-T2-02_Diplura 0.257 0.257 0.197 0.252 0.254 0.242 0.172 0.277 0.258 0.223 0.232 0.238 0.161 0.152 0.221 0.230 0.249 0.260 0.266 0.252 0.237 0.163 0.158 0.155 0.169 0.164 0.210 0.213 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 24. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV167 IV171 IV172 IV175 IV176 IV177 IV178 IV179 IV181 IV189 Anas Bsp Hsp10 Hsp12 Hsp21 Lyal Psp1 Psp4 B12 EO10 EO11 EO12 J17-2 G173 G175 G350 G92 J17-5 J17-9 J17-11 I49 IE37 IE39 IE40 IE45 IE50 IS105 IS111 IS126 IS92 IS97 IT40 IV167_M2ERC053-T2-01_Isopoda 0.016 0.016 0.009 0.016 0.016 0.016 0.015 0.016 0.015 0.017 0.016 0.014 0.015 0.014 0.015 0.017 0.016 0.016 0.016 0.016 0.018 0.018 0.013 0.015 0.016 0.017 0.017 0.016 0.015 0.020 0.016 0.022 0.017 0.020 0.018 0.017 0.020 0.013 0.018 0.024 0.013 IV171_MEARC4923-20150930-T1-01_Isopoda 0.225 0.014 0.014 0.016 0.014 0.016 0.015 0.007 0.015 0.015 0.016 0.015 0.015 0.014 0.016 0.016 0.018 0.017 0.015 0.013 0.014 0.016 0.015 0.014 0.013 0.014 0.015 0.015 0.016 0.021 0.014 0.025 0.014 0.010 0.021 0.017 0.017 0.015 0.015 0.024 0.015 IV172_MELUNK02-20151001-T1-03_Isopoda 0.203 0.238 0.015 0.017 0.015 0.016 0.015 0.015 0.015 0.015 0.016 0.016 0.018 0.015 0.015 0.018 0.018 0.016 0.015 0.014 0.015 0.016 0.014 0.014 0.014 0.016 0.017 0.016 0.017 0.022 0.016 0.020 0.015 0.019 0.018 0.016 0.020 0.015 0.016 0.021 0.015 IV175_RC13MRR0014-20151002-T1-01_Isopoda 0.070 0.224 0.191 0.016 0.015 0.015 0.016 0.014 0.014 0.016 0.015 0.014 0.016 0.015 0.015 0.016 0.017 0.017 0.015 0.014 0.016 0.018 0.013 0.016 0.017 0.017 0.016 0.016 0.014 0.019 0.015 0.022 0.015 0.018 0.018 0.016 0.019 0.013 0.017 0.021 0.012 IV176_RC14MEB0060-20151001-T2-03_Isopoda 0.240 0.231 0.259 0.242 0.017 0.017 0.011 0.016 0.011 0.016 0.016 0.016 0.015 0.016 0.017 0.019 0.020 0.016 0.016 0.016 0.015 0.014 0.017 0.016 0.015 0.014 0.016 0.017 0.017 0.018 0.015 0.023 0.015 0.020 0.021 0.015 0.021 0.016 0.016 0.022 0.014 IV177_RC14MEB0088-20151001-T3-01_Isopoda 0.227 0.166 0.226 0.220 0.218 0.017 0.016 0.015 0.017 0.014 0.016 0.014 0.015 0.014 0.016 0.016 0.017 0.017 0.016 0.014 0.015 0.016 0.014 0.014 0.014 0.016 0.016 0.015 0.015 0.020 0.016 0.023 0.014 0.019 0.019 0.018 0.020 0.017 0.017 0.024 0.016 IV178_RC14MEB0101-20151001-T1-01_Isopoda 0.189 0.201 0.194 0.191 0.224 0.238 0.017 0.016 0.015 0.016 0.016 0.016 0.017 0.014 0.016 0.018 0.019 0.018 0.016 0.016 0.017 0.016 0.016 0.016 0.016 0.016 0.017 0.017 0.017 0.023 0.015 0.021 0.015 0.019 0.021 0.017 0.018 0.015 0.017 0.022 0.014 IV179_TOBRC0020-20151001-T2-02_Isopoda 0.240 0.201 0.238 0.232 0.082 0.207 0.227 0.015 0.010 0.014 0.016 0.015 0.013 0.016 0.016 0.019 0.018 0.017 0.016 0.014 0.015 0.016 0.016 0.015 0.015 0.014 0.015 0.017 0.017 0.019 0.014 0.023 0.015 0.020 0.020 0.015 0.019 0.015 0.015 0.022 0.016 IV181_MEARC2401-20150807-T2-03_Isopoda 0.230 0.038 0.230 0.224 0.228 0.169 0.207 0.207 0.015 0.016 0.016 0.016 0.015 0.015 0.016 0.017 0.018 0.017 0.014 0.014 0.015 0.016 0.016 0.015 0.014 0.015 0.016 0.015 0.016 0.019 0.016 0.024 0.015 0.006 0.022 0.017 0.018 0.016 0.017 0.024 0.016 IV189_TOBRC0020-t1-01_Isopoda 0.241 0.216 0.242 0.236 0.076 0.211 0.235 0.067 0.207 0.014 0.016 0.014 0.014 0.015 0.016 0.020 0.017 0.016 0.014 0.014 0.015 0.015 0.015 0.015 0.014 0.015 0.014 0.017 0.016 0.019 0.014 0.022 0.014 0.020 0.018 0.016 0.020 0.014 0.015 0.022 0.014 FN824099_Armadillidium_nasatum 0.203 0.215 0.195 0.206 0.223 0.219 0.189 0.212 0.212 0.212 0.015 0.016 0.015 0.016 0.014 0.017 0.016 0.016 0.016 0.015 0.016 0.016 0.016 0.014 0.015 0.016 0.016 0.018 0.017 0.020 0.015 0.023 0.016 0.020 0.017 0.016 0.018 0.015 0.016 0.020 0.015 AB626254_Burmoniscus_sp. 0.222 0.210 0.244 0.210 0.229 0.206 0.207 0.209 0.216 0.219 0.206 0.015 0.016 0.015 0.016 0.019 0.019 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.016 0.018 0.017 0.021 0.017 0.021 0.015 0.020 0.017 0.017 0.020 0.017 0.018 0.022 0.016 Haloniscus_sp._10_EU364592 0.193 0.230 0.197 0.194 0.232 0.219 0.196 0.227 0.228 0.230 0.182 0.205 0.013 0.013 0.016 0.017 0.017 0.017 0.016 0.016 0.017 0.017 0.015 0.014 0.016 0.015 0.016 0.017 0.018 0.019 0.017 0.021 0.017 0.018 0.018 0.017 0.019 0.015 0.016 0.022 0.014 Haloniscus_sp._12_EU364601 0.206 0.233 0.230 0.223 0.227 0.231 0.208 0.220 0.231 0.222 0.197 0.220 0.149 0.014 0.014 0.017 0.016 0.016 0.015 0.014 0.015 0.017 0.015 0.012 0.017 0.014 0.015 0.015 0.017 0.019 0.015 0.023 0.016 0.019 0.019 0.016 0.021 0.016 0.015 0.022 0.017 EU364622_Haloniscus_sp._21 0.186 0.202 0.213 0.190 0.225 0.217 0.191 0.213 0.208 0.211 0.204 0.222 0.147 0.168 0.013 0.016 0.016 0.017 0.015 0.016 0.015 0.016 0.013 0.013 0.015 0.013 0.015 0.016 0.013 0.019 0.015 0.020 0.016 0.018 0.016 0.016 0.020 0.014 0.016 0.022 0.015 EU364629_Laevophiloscia_yalgoorensis 0.193 0.250 0.202 0.197 0.235 0.246 0.212 0.241 0.246 0.239 0.210 0.230 0.207 0.223 0.221 0.017 0.016 0.015 0.017 0.013 0.017 0.016 0.015 0.015 0.015 0.015 0.015 0.015 0.016 0.020 0.015 0.022 0.016 0.021 0.018 0.017 0.022 0.014 0.019 0.019 0.014 EU107646_Pygolabis_sp._1 0.248 0.259 0.264 0.239 0.282 0.250 0.253 0.255 0.260 0.282 0.244 0.240 0.240 0.264 0.235 0.257 0.014 0.019 0.018 0.016 0.019 0.020 0.017 0.017 0.018 0.017 0.017 0.018 0.019 0.018 0.017 0.023 0.019 0.022 0.018 0.019 0.024 0.018 0.018 0.025 0.016 EU107664_Pygolabis_sp._4 0.224 0.253 0.246 0.228 0.275 0.266 0.250 0.270 0.250 0.282 0.220 0.240 0.228 0.240 0.242 0.240 0.155 0.020 0.018 0.017 0.018 0.019 0.017 0.017 0.019 0.018 0.016 0.017 0.018 0.017 0.018 0.024 0.018 0.020 0.020 0.019 0.023 0.017 0.019 0.023 0.015 b12_20_1b_Isopoda 0.269 0.292 0.292 0.269 0.290 0.279 0.287 0.292 0.289 0.295 0.255 0.266 0.243 0.236 0.266 0.249 0.279 0.275 0.017 0.016 0.018 0.019 0.016 0.015 0.018 0.019 0.017 0.017 0.018 0.022 0.017 0.024 0.018 0.022 0.022 0.018 0.023 0.016 0.019 0.023 0.018 EO10_Troglarmadillo_sp._B39_MG0198R 0.232 0.204 0.211 0.230 0.215 0.200 0.239 0.198 0.198 0.201 0.209 0.221 0.224 0.211 0.217 0.230 0.251 0.253 0.284 0.014 0.015 0.016 0.015 0.017 0.015 0.014 0.014 0.015 0.017 0.020 0.014 0.020 0.015 0.018 0.019 0.015 0.021 0.018 0.015 0.022 0.017 EO11_Troglarmadillo_ISO005_EKP0042 0.217 0.219 0.220 0.213 0.250 0.223 0.227 0.227 0.219 0.227 0.230 0.227 0.227 0.239 0.211 0.227 0.268 0.253 0.292 0.197 0.014 0.017 0.016 0.014 0.013 0.014 0.014 0.013 0.014 0.021 0.012 0.021 0.013 0.018 0.020 0.015 0.018 0.016 0.014 0.021 0.016 EO12_Troglarmadillo_sp._B42_EEX0561 0.246 0.224 0.235 0.249 0.198 0.232 0.236 0.181 0.219 0.190 0.235 0.233 0.244 0.236 0.231 0.258 0.251 0.244 0.326 0.190 0.210 0.014 0.016 0.017 0.013 0.012 0.015 0.014 0.014 0.022 0.014 0.020 0.016 0.019 0.020 0.015 0.020 0.018 0.015 0.021 0.018 EXR1356_J17-2 0.243 0.237 0.243 0.246 0.209 0.247 0.245 0.189 0.235 0.205 0.227 0.246 0.244 0.257 0.236 0.247 0.284 0.253 0.311 0.209 0.229 0.174 0.017 0.017 0.014 0.016 0.014 0.016 0.016 0.020 0.014 0.020 0.015 0.020 0.023 0.016 0.019 0.018 0.015 0.023 0.016 G173_Philosciidae_S1_100421 0.188 0.210 0.214 0.193 0.237 0.224 0.204 0.231 0.216 0.225 0.200 0.214 0.163 0.189 0.156 0.210 0.242 0.237 0.270 0.221 0.222 0.247 0.248 0.014 0.014 0.016 0.015 0.017 0.015 0.017 0.016 0.021 0.016 0.021 0.017 0.016 0.020 0.016 0.015 0.022 0.014 G175_Philosciidae_S1_LN7657 0.193 0.228 0.216 0.191 0.237 0.231 0.224 0.224 0.239 0.239 0.193 0.227 0.161 0.170 0.160 0.207 0.246 0.240 0.256 0.227 0.230 0.234 0.245 0.178 0.014 0.015 0.015 0.016 0.015 0.020 0.015 0.021 0.015 0.020 0.016 0.014 0.019 0.014 0.015 0.022 0.015 G350_LN9740_Troglamadillo_sp_OP 0.225 0.225 0.229 0.219 0.188 0.220 0.223 0.200 0.216 0.197 0.210 0.230 0.219 0.217 0.210 0.238 0.266 0.240 0.311 0.198 0.204 0.145 0.160 0.222 0.229 0.015 0.015 0.017 0.015 0.020 0.015 0.019 0.014 0.017 0.023 0.016 0.020 0.018 0.016 0.023 0.017 G92_Troglarmadillo_sp_LN8501 0.230 0.197 0.242 0.239 0.201 0.204 0.216 0.181 0.193 0.185 0.234 0.214 0.227 0.228 0.213 0.238 0.259 0.266 0.316 0.196 0.201 0.182 0.217 0.236 0.233 0.204 0.015 0.014 0.016 0.019 0.014 0.020 0.016 0.019 0.020 0.015 0.020 0.017 0.013 0.023 0.016 GA0136R_J17-5 0.217 0.206 0.234 0.217 0.236 0.217 0.220 0.216 0.205 0.214 0.229 0.209 0.203 0.222 0.205 0.228 0.273 0.253 0.282 0.196 0.141 0.203 0.217 0.208 0.233 0.193 0.183 0.014 0.013 0.020 0.014 0.017 0.014 0.020 0.019 0.016 0.018 0.018 0.014 0.020 0.015 GFR002_J17-9 0.246 0.221 0.243 0.238 0.220 0.226 0.236 0.211 0.218 0.206 0.252 0.229 0.228 0.238 0.219 0.241 0.264 0.260 0.297 0.203 0.168 0.209 0.217 0.245 0.243 0.221 0.194 0.148 0.012 0.021 0.014 0.019 0.015 0.020 0.022 0.017 0.021 0.018 0.015 0.020 0.016 GSR0020_J17-11 0.238 0.215 0.247 0.232 0.234 0.224 0.251 0.220 0.215 0.224 0.251 0.224 0.230 0.233 0.217 0.247 0.273 0.264 0.289 0.218 0.174 0.235 0.231 0.245 0.253 0.223 0.208 0.150 0.130 0.021 0.014 0.019 0.015 0.021 0.020 0.016 0.020 0.017 0.016 0.021 0.015 I49-PT04_Isopoda 0.258 0.274 0.263 0.247 0.291 0.271 0.289 0.284 0.267 0.295 0.256 0.256 0.256 0.263 0.271 0.293 0.171 0.155 0.265 0.269 0.289 0.295 0.306 0.265 0.289 0.278 0.287 0.269 0.280 0.274 0.021 0.027 0.019 0.024 0.022 0.022 0.026 0.019 0.020 0.026 0.018 IE37_BHRC122-20141216-T2-03 0.222 0.223 0.233 0.220 0.218 0.214 0.213 0.195 0.217 0.202 0.227 0.230 0.218 0.223 0.214 0.236 0.271 0.259 0.284 0.196 0.170 0.204 0.208 0.228 0.232 0.209 0.191 0.167 0.162 0.166 0.282 0.018 0.015 0.019 0.020 0.015 0.018 0.016 0.015 0.021 0.015 IE39_TBRC045P3T1-3 0.246 0.267 0.270 0.270 0.219 0.258 0.227 0.217 0.267 0.229 0.262 0.262 0.264 0.257 0.245 0.266 0.313 0.288 0.317 0.253 0.251 0.243 0.248 0.248 0.281 0.236 0.243 0.222 0.262 0.258 0.318 0.239 0.024 0.023 0.023 0.024 0.021 0.022 0.021 0.025 0.022 IE40_CWRC155P3T2-1 0.222 0.214 0.217 0.224 0.223 0.218 0.230 0.204 0.217 0.205 0.221 0.200 0.228 0.227 0.226 0.213 0.271 0.253 0.280 0.192 0.214 0.211 0.212 0.220 0.232 0.204 0.207 0.222 0.227 0.217 0.267 0.218 0.243 0.019 0.018 0.018 0.021 0.017 0.017 0.022 0.015 IE45_MEARC2999P6T2-1 0.231 0.038 0.261 0.232 0.227 0.169 0.223 0.218 0.012 0.228 0.223 0.223 0.236 0.248 0.224 0.250 0.270 0.260 0.294 0.216 0.244 0.239 0.254 0.225 0.254 0.232 0.221 0.223 0.232 0.225 0.259 0.235 0.267 0.230 0.023 0.021 0.019 0.019 0.019 0.024 0.017 IE50_RC08MEJ0076P2T2-1 0.297 0.296 0.301 0.272 0.283 0.275 0.288 0.283 0.294 0.272 0.268 0.275 0.222 0.248 0.233 0.296 0.321 0.321 0.274 0.281 0.305 0.301 0.312 0.248 0.257 0.277 0.309 0.268 0.305 0.327 0.323 0.296 0.316 0.303 0.306 0.020 0.024 0.016 0.021 0.023 0.018 IS105_TSOOPP03_20150709_T2_03_Lin_3 0.262 0.216 0.236 0.256 0.230 0.232 0.244 0.218 0.220 0.226 0.242 0.244 0.236 0.245 0.248 0.264 0.266 0.273 0.291 0.231 0.226 0.245 0.237 0.255 0.241 0.235 0.201 0.198 0.213 0.213 0.278 0.216 0.265 0.258 0.232 0.294 0.021 0.017 0.014 0.022 0.018 IS111_CWRC937_20150912_T2_01_Lin_2 0.237 0.215 0.223 0.228 0.221 0.228 0.225 0.220 0.212 0.223 0.230 0.238 0.223 0.247 0.244 0.262 0.299 0.274 0.309 0.217 0.230 0.240 0.246 0.253 0.246 0.233 0.233 0.217 0.228 0.238 0.305 0.205 0.240 0.253 0.223 0.302 0.240 0.019 0.016 0.021 0.021 IS126_KBRC1154_20150909_T2_01_Lin_6 0.154 0.229 0.183 0.148 0.240 0.243 0.183 0.229 0.235 0.235 0.188 0.218 0.191 0.209 0.195 0.195 0.242 0.231 0.276 0.215 0.232 0.240 0.250 0.202 0.212 0.240 0.239 0.240 0.242 0.237 0.256 0.232 0.258 0.238 0.246 0.292 0.252 0.261 0.016 0.024 0.014 IS92_RHRC414_20150709_T2_02_Lin_1 0.240 0.206 0.233 0.242 0.211 0.203 0.231 0.198 0.209 0.203 0.237 0.234 0.233 0.225 0.211 0.251 0.275 0.284 0.291 0.203 0.209 0.220 0.250 0.255 0.233 0.221 0.190 0.186 0.202 0.204 0.293 0.194 0.241 0.232 0.235 0.277 0.215 0.120 0.253 0.022 0.018 IS97_CWRC485.20150709_T1_04_Lin_5 0.234 0.251 0.251 0.227 0.256 0.264 0.199 0.258 0.256 0.264 0.240 0.251 0.223 0.220 0.252 0.205 0.311 0.291 0.256 0.258 0.251 0.264 0.276 0.227 0.251 0.261 0.256 0.227 0.269 0.266 0.322 0.253 0.279 0.230 0.258 0.287 0.276 0.238 0.253 0.240 0.023 IT40_BHRCOPP1-151013-T3-02_Isopoda 0.130 0.228 0.193 0.121 0.227 0.229 0.196 0.223 0.231 0.220 0.209 0.206 0.200 0.201 0.191 0.186 0.244 0.222 0.272 0.223 0.223 0.261 0.236 0.184 0.204 0.226 0.234 0.217 0.237 0.234 0.245 0.229 0.272 0.228 0.232 0.288 0.264 0.246 0.140 0.239 0.233 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 29. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV212 IV226 IV203 IV210 Aussp Tyrsp Lagjo CM18 G319 G324 G328 G219 G220 G118 J20-6 PI012 EA0270R LB084 EAP0178 GF0173R JIN0078R EJR0202 IS01 IS05 IS06 IS08 IS10 IS11 IS12 IV212_DD14MEL0001-20151001-T2-02_lin_9 0.015 0.018 0.019 0.019 0.022 0.022 0.019 0.023 0.019 0.019 0.019 0.021 0.024 0.024 0.020 0.025 0.022 0.022 0.022 0.024 0.024 0.018 0.020 0.018 0.019 0.021 0.017 0.020 IV226_RC14MRR0020-20151002-T2-01_lin_10 0.111 0.018 0.019 0.019 0.021 0.021 0.018 0.021 0.021 0.018 0.020 0.023 0.022 0.021 0.022 0.024 0.021 0.020 0.020 0.022 0.023 0.018 0.020 0.019 0.019 0.020 0.016 0.019 IV203_DD14MRR0004T1-04_lin_11 0.204 0.207 0.017 0.018 0.021 0.017 0.016 0.020 0.017 0.019 0.016 0.017 0.022 0.021 0.020 0.022 0.022 0.023 0.020 0.022 0.021 0.019 0.018 0.017 0.017 0.018 0.015 0.019 IV210_MEARC3814-20150606-02sc_lin_12 0.218 0.238 0.156 0.019 0.023 0.022 0.019 0.020 0.019 0.018 0.019 0.020 0.020 0.022 0.022 0.024 0.023 0.021 0.021 0.020 0.020 0.016 0.018 0.016 0.016 0.017 0.019 0.019 EU559513_Austrochtonia_sp 0.196 0.207 0.166 0.189 0.023 0.019 0.018 0.019 0.017 0.020 0.019 0.020 0.023 0.024 0.022 0.025 0.023 0.025 0.023 0.023 0.023 0.020 0.019 0.020 0.021 0.020 0.018 0.021 EU559506_Tyrannochthonius_sp._JM-2008 0.296 0.293 0.283 0.278 0.261 0.023 0.020 0.021 0.020 0.021 0.021 0.021 0.022 0.000 0.022 0.021 0.000 0.024 0.021 0.022 0.022 0.022 0.023 0.023 0.024 0.024 0.021 0.023 EU559503_Lagynochthonius_johni 0.231 0.226 0.215 0.233 0.203 0.276 0.021 0.020 0.020 0.020 0.022 0.020 0.024 0.023 0.020 0.024 0.023 0.025 0.022 0.024 0.023 0.022 0.021 0.021 0.020 0.020 0.019 0.021 CM18_RC11KOOD0193P2T1-2_Lagynochthonius_sp. 0.216 0.216 0.182 0.216 0.203 0.254 0.226 0.019 0.017 0.018 0.017 0.019 0.022 0.021 0.024 0.024 0.020 0.026 0.018 0.022 0.021 0.019 0.018 0.018 0.019 0.018 0.018 0.020 G319_LN8720_Chthoniid_sp_MJ 0.247 0.249 0.210 0.220 0.191 0.289 0.235 0.242 0.018 0.019 0.020 0.020 0.020 0.022 0.022 0.023 0.021 0.025 0.022 0.020 0.020 0.024 0.021 0.021 0.021 0.019 0.020 0.024 G324_101086_Tyrannochthonius_sp_MA 0.212 0.212 0.176 0.203 0.179 0.252 0.220 0.178 0.191 0.017 0.018 0.019 0.022 0.019 0.023 0.021 0.020 0.024 0.019 0.022 0.021 0.019 0.018 0.019 0.019 0.017 0.017 0.020 G328_LN9199_Tyrannochthonius_sp_OP 0.211 0.216 0.187 0.199 0.181 0.208 0.236 0.180 0.209 0.171 0.020 0.018 0.022 0.020 0.021 0.022 0.021 0.022 0.018 0.022 0.021 0.019 0.019 0.019 0.020 0.021 0.018 0.020 G219_Pseudoscorpian_108935 0.216 0.221 0.184 0.179 0.176 0.280 0.226 0.198 0.225 0.187 0.169 0.014 0.022 0.021 0.022 0.024 0.020 0.024 0.023 0.022 0.022 0.016 0.017 0.016 0.018 0.020 0.015 0.017 G220_Pseudoscorpian_109177 0.211 0.221 0.177 0.195 0.185 0.293 0.206 0.202 0.237 0.191 0.184 0.117 0.022 0.021 0.022 0.021 0.021 0.025 0.022 0.022 0.020 0.017 0.018 0.017 0.018 0.020 0.019 0.018 G118_Tyrannochthonius 0.275 0.278 0.238 0.238 0.238 0.240 0.261 0.278 0.241 0.252 0.249 0.232 0.235 0.022 0.018 0.022 0.022 0.024 0.022 0.000 0.000 0.022 0.025 0.024 0.023 0.023 0.021 0.021 EXR1148_J20-6_Ministers_N_Lagy_S4 0.234 0.234 0.216 0.207 0.201 0.000 0.216 0.192 0.243 0.201 0.178 0.216 0.225 0.231 0.022 0.021 0.000 0.024 0.021 0.022 0.022 0.022 0.023 0.023 0.024 0.024 0.022 0.022 PI012_Hashimoto_Tyran 0.216 0.246 0.183 0.207 0.192 0.197 0.222 0.210 0.210 0.216 0.160 0.189 0.192 0.195 0.197 0.021 0.022 0.023 0.021 0.018 0.018 0.023 0.024 0.022 0.023 0.023 0.019 0.025 EA0270R_OB24_Lag_B2 0.265 0.268 0.214 0.223 0.229 0.228 0.265 0.232 0.271 0.193 0.202 0.193 0.199 0.265 0.228 0.228 0.021 0.024 0.021 0.022 0.022 0.023 0.021 0.021 0.022 0.023 0.021 0.023 LB084_Hashimoto_Tyrann 0.291 0.288 0.277 0.272 0.259 0.000 0.275 0.246 0.289 0.252 0.208 0.275 0.288 0.240 0.000 0.197 0.228 0.024 0.021 0.022 0.022 0.022 0.023 0.023 0.025 0.024 0.021 0.023 EAP0178_OB24_Tyran_B1 0.218 0.221 0.253 0.250 0.241 0.267 0.282 0.268 0.279 0.247 0.226 0.235 0.268 0.291 0.267 0.267 0.280 0.267 0.019 0.024 0.024 0.022 0.023 0.025 0.022 0.023 0.022 0.022 GF0173R_MAC_dep_F_Lag_B2 0.255 0.249 0.208 0.235 0.194 0.189 0.223 0.188 0.223 0.188 0.155 0.205 0.211 0.226 0.189 0.183 0.238 0.189 0.265 0.022 0.022 0.022 0.024 0.022 0.022 0.023 0.020 0.022 JIN0078R_BHP772_Jinaryi_Lagynochthonius 0.277 0.283 0.241 0.241 0.241 0.240 0.263 0.283 0.244 0.255 0.252 0.235 0.238 0.000 0.231 0.195 0.265 0.240 0.291 0.226 0.000 0.022 0.025 0.024 0.023 0.023 0.021 0.022 EJR0202_2_9_9_ob18_Tyrranochthonius_B2 0.252 0.255 0.217 0.217 0.217 0.231 0.243 0.261 0.223 0.232 0.226 0.214 0.214 0.000 0.231 0.195 0.265 0.231 0.291 0.226 0.000 0.021 0.025 0.024 0.023 0.021 0.020 0.021 IS01 0.187 0.207 0.204 0.198 0.214 0.308 0.249 0.238 0.230 0.210 0.206 0.198 0.211 0.306 0.240 0.234 0.256 0.304 0.206 0.258 0.311 0.287 0.017 0.016 0.012 0.019 0.020 0.008 IS05 0.204 0.202 0.236 0.222 0.244 0.303 0.242 0.220 0.267 0.215 0.225 0.168 0.200 0.295 0.246 0.263 0.259 0.298 0.224 0.264 0.300 0.276 0.167 0.014 0.018 0.018 0.018 0.016 IS06 0.187 0.189 0.193 0.216 0.228 0.311 0.229 0.222 0.249 0.217 0.206 0.177 0.200 0.306 0.249 0.246 0.256 0.306 0.238 0.273 0.311 0.284 0.149 0.116 0.017 0.019 0.020 0.015 IS08 0.189 0.216 0.216 0.207 0.230 0.311 0.256 0.236 0.237 0.208 0.230 0.205 0.214 0.309 0.246 0.260 0.271 0.306 0.221 0.267 0.311 0.290 0.093 0.149 0.136 0.018 0.020 0.013 IS10 0.240 0.238 0.200 0.193 0.210 0.298 0.229 0.224 0.235 0.215 0.221 0.209 0.230 0.255 0.228 0.228 0.250 0.293 0.268 0.238 0.258 0.238 0.227 0.231 0.258 0.222 0.019 0.019 IS11 0.193 0.193 0.144 0.187 0.184 0.285 0.210 0.189 0.237 0.187 0.194 0.166 0.189 0.235 0.225 0.178 0.244 0.280 0.294 0.194 0.238 0.214 0.229 0.209 0.227 0.222 0.176 0.022 IS12 0.196 0.202 0.204 0.204 0.217 0.303 0.249 0.220 0.232 0.203 0.199 0.186 0.195 0.309 0.240 0.243 0.247 0.296 0.197 0.249 0.314 0.290 0.053 0.147 0.147 0.091 0.236 0.233 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 30. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV201 IV217 IV206 IV209 IV202 IV211 IV213 IV219 IV205 IV208 IV207 Indsp DF18 FZ1 G107 GB0018R GC0103R IT72 PSF0090R Parsu IV201_DD14MEB0002-20150607-01sc_Pseudoscorpiones 0.009 0.018 0.018 0.022 0.021 0.022 0.021 0.021 0.018 0.020 0.020 0.020 0.021 0.023 0.025 0.025 0.023 0.024 0.021 IV217_RC14MEB0029-20151001-T1-01_Pseudoscorpiones 0.049 0.020 0.019 0.023 0.022 0.021 0.022 0.022 0.020 0.021 0.020 0.020 0.023 0.024 0.025 0.025 0.025 0.025 0.021 IV206_DD14MRR0008-20150808-T1-02_Pseudoscorpiones 0.213 0.227 0.009 0.020 0.019 0.022 0.020 0.018 0.019 0.019 0.019 0.018 0.020 0.021 0.025 0.024 0.022 0.024 0.020 IV209_M2ERC027-t2-03_Pseudoscorpiones 0.207 0.222 0.036 0.020 0.018 0.021 0.020 0.018 0.018 0.019 0.020 0.018 0.020 0.021 0.025 0.024 0.023 0.023 0.020 IV202_DD14MRR0004T1-04_Pseudoscorpiones 0.267 0.260 0.262 0.262 0.022 0.022 0.021 0.021 0.022 0.022 0.022 0.020 0.022 0.026 0.029 0.028 0.023 0.028 0.022 IV211_RC13MEA0279-20150807-T1-02_Pseudoscorpiones 0.227 0.224 0.240 0.227 0.224 0.018 0.018 0.022 0.019 0.021 0.020 0.022 0.021 0.025 0.028 0.028 0.020 0.027 0.020 IV213_DD14MRR0009-20151002-T2-02_Pseudoscorpiones 0.253 0.258 0.249 0.240 0.227 0.131 0.017 0.023 0.020 0.021 0.018 0.021 0.020 0.026 0.030 0.030 0.017 0.028 0.023 IV219_RC14MEB0101-20151001-T1-02_Pseudoscorpiones 0.252 0.254 0.261 0.252 0.242 0.141 0.134 0.022 0.020 0.020 0.020 0.024 0.018 0.025 0.027 0.027 0.016 0.026 0.020 IV205_DD14MRR0005-T2-01_Pseudoscorpiones 0.298 0.304 0.309 0.304 0.282 0.282 0.287 0.289 0.017 0.020 0.021 0.021 0.023 0.025 0.027 0.026 0.023 0.026 0.019 IV208_M2ERC027-t2-03_Pseudoscorpiones 0.264 0.276 0.284 0.276 0.284 0.253 0.256 0.256 0.136 0.012 0.019 0.021 0.022 0.023 0.025 0.025 0.021 0.024 0.019 IV207_DD14MRR0008-20150808-T2-03_Pseudoscorpiones 0.280 0.296 0.313 0.296 0.296 0.260 0.264 0.275 0.167 0.082 0.019 0.021 0.023 0.025 0.026 0.025 0.023 0.025 0.019 EU559564_Indohya_sp._JM-2008 0.211 0.211 0.216 0.211 0.247 0.216 0.229 0.219 0.273 0.244 0.260 0.021 0.022 0.025 0.029 0.027 0.019 0.026 0.020 DF18_8668_Indohya_sp. 0.216 0.218 0.162 0.162 0.236 0.218 0.220 0.254 0.293 0.264 0.282 0.211 0.020 0.022 0.027 0.029 0.021 0.026 0.020 FZ1_Pseudoscorpion_RC13KOOD0302P7T1-1 0.224 0.233 0.231 0.228 0.264 0.221 0.228 0.215 0.296 0.276 0.296 0.228 0.200 0.023 0.024 0.023 0.022 0.025 0.022 G107_Indohya 0.258 0.255 0.176 0.167 0.284 0.284 0.308 0.282 0.340 0.314 0.340 0.252 0.199 0.258 0.028 0.026 0.024 0.024 0.025 GB0018R_Hyidae_B1 0.252 0.255 0.264 0.261 0.337 0.290 0.305 0.305 0.349 0.349 0.361 0.293 0.255 0.238 0.276 0.012 0.028 0.022 0.027 GC0103R_Hyidae 0.255 0.255 0.246 0.249 0.326 0.293 0.302 0.296 0.346 0.343 0.361 0.276 0.258 0.235 0.258 0.053 0.028 0.024 0.027 IT72_BHRC0512-151013-T3-01_Pseudoscorpiones 0.245 0.247 0.263 0.263 0.231 0.113 0.105 0.113 0.285 0.258 0.272 0.191 0.215 0.230 0.268 0.287 0.277 0.027 0.022 PSF0090R_LN2124_Indohya 0.273 0.279 0.258 0.258 0.349 0.311 0.299 0.323 0.378 0.358 0.361 0.287 0.282 0.276 0.287 0.173 0.176 0.283 0.028 JN018204_Paratemnoides_sumatranus_Atemnidae 0.289 0.298 0.287 0.280 0.276 0.267 0.267 0.282 0.184 0.180 0.198 0.277 0.277 0.308 0.337 0.337 0.323 0.272 0.352 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. Disclaimer:The number Although of base utmost differences care has been per taken site to fromensure between the correctness sequences of the caption, are shown. the caption Standard text is provided error "asestimate(s) is" without any are warranty shown of above any kind. the Authors diagonal. advise Thethe user analysis to carefully involved check the 42 caption nucleotide prior to sequences.its use for any purposeCodon and positions report any included errors or were problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application.

Molecular systematics of subfauna from the Robe Valley

27 September, 2016

Jason Alexander Biota Environmental Sciences Level 1, 228 Carr Place Leederville, WA 6007

Via email

Re. Report on the molecular systematics of subfauna from the Robe River Valley – Phase 3 and historical samples – Part 1.

Dear Jason,

Following is a summary of the results of the subfauna study we have completed on four of the 11 taxonomic groups from the Robe River Valley. Thirty-one distinct genetic lineages were detected among the four groups so far analysed. The 31 lineages likely represent between 24 and 30 species, up to 11 of which may have been detected previously in the Pilbara. Further investigation may be required to resolve relationships between closely related lineages.

Sincerely,

Dr. Terrie Finston, Yvette Hitchen and Dr. Oliver Berry Helix Molecular Solutions

1 Molecular systematics of subfauna from the Robe Valley

Background and Objective Seventy-two specimens of subfauna belonging to four taxonomic groups (Araneae, Pseudoscorpiones, Schizomida, Thysanura) were collected from Robe River Valley and sequenced for variation at the mitochondrial COI gene. An additional seven taxonomic groups are still to be analysed and will be covered in a separate report. The molecular data were assessed in order to determine the number of species present in each group and compare the results to those obtained during previous surveys that have been undertaken for the these groups elsewhere in the Pilbara.

Executive summary  Seventy-two specimens from the Robe River Valley belonging to four taxonomic groups were sequenced for variation at the COI gene or 12s when necessary.  31 lineages were detected among the four groups.  The 31 lineages likely represent between 24 and 30 species.  Up to 11 of the species may have been detected previously in the Pilbara, whereas the remainder appear to be new, based on the material available for comparison, however, further investigation may be required to resolve relationships between closely related lineages.

Methods Seventy-two specimens of subfauna collected from the Robe River Valley area were sequenced for variation at the mitochondrial cytochrome oxidase subunit I gene (COI) using multiple pairs of primers (LCOI/HCO2, LCOI-long/HCO2-long, NemF1/NemR1 and LCOI/CIN2341. The 12s region was amplified and sequenced using primers 12-ai and 12s-bi (Simon et al., 1994).

Results Araneae Seven historical specimens of Araneae were sequenced for COI (Table 1). Sequences were obtained from five specimens and these were analysed with 24 lineages of Araneae from the Pilbara, representing 36 specimens, as well as five GenBank reference specimens as follows: Aranidae (Arachnura scorpiooides, GenBank accession #KJ957946 and Gea theridoides, Genbank accession #KJ959766), Oonopidae (Ischnothyreus auritus, Genbank accession# KR864743) Theridiidae (Theridion ohlerti Genbank accession #GU684645 and Robertus neglectus GenBank accession #AY231053).

2 Molecular systematics of subfauna from the Robe Valley

Phylogenetic analysis The phylogenetic analysis, which included the five specimens from the Robe Valley in addition to 29 reference specimens, placed the Robe Valley specimens in four genetically distinct clades (Figure 1). Two of the lineages were placed in a large clade containing reference specimens of Oonopidae and were assigned to the new lineages AO014 and AO015 (Figure 1). In particular, AO015 formed a well-supported clade with the Genbank specimen of Oonopidae; Figure 1). The third lineage (AA001) was placed in a well-supported clade containing the two Genbank specimens of Aranidae (Figure 1) and the fourth lineage (A012) was placed in a clade containing Pilbara specimens assigned to the genus Anapistula (family Symphytognathidae) on the basis of morphology, however this clade formed a well-supported clade with the Genbank reference specimens of Theriidae (Figure 1).

Differentiation within and between lineages The Oonopidae lineage AO014 differed from the reference specimens by between 14.1 and 25.1% sequence divergence and Oonopidae lineage AO015 differed from the reference specimens by between 16.4 and 24.8% sequence divergence (Table 2). The Aranidae lineage AA001 differed from the reference lineages by between 11.4 and 30.3% sequence divergence and the final lineage, A012, differed from the reference lineages by between 10.8 and 31.4% (Table 2).

Pseudoscorpions Reference sequences and outgroups Twenty-seven pseudoscorpions (historic, phase 3 and repeats from phases 1 and 2) were sequenced from the Robe Valley (Table 3). In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted on the 22 Robe Valley pseudoscorpions for which sequences were obtained. Fourteen distinct genetic lineages were detected and haplotypes were selected from each genetic lineage to include in a model-based phylogenetic analysis. A search of similar sequences on Genbank and in the Helix database indicated that five of the lineages (1 – 5) most closely matched the family Chthoniidae, seven of the lineages (8 –14) most closely matched reference specimens of Hyiidae and two of the lineages (6 and 7) most closely matched reference specimens of Olpiidae. Based on those results, two separate phylogenetic analyses were conducted, one including the Chthoniidae and one including both the Hyiidae and the Olpiidae. Genbank voucher sequences of Chthoniidae were included in the former analysis as follows: nine sequences belonging to eight genera of Chthoniidae: Anaulacodithella sp (Genbank accession # EU559520), Sathrochthonius insulanus (Genbank accession #EU559521), Pseudochthonius sp. (Genbank accession #EU559519), Pseudotyrannochthonius s. JM2 (Genbank accession #EU559509), Paraliochthonius sp JM 2008 (Genbank accession #EU5595505), Tyrannochthonius sp JM 2008 (Genbank accession # EU559506) from South America (Columbia), Tyrannochthonius aridus (GenBank accession # KJ659959 from Western Australia), Lagynochthonius johni (Genbank accession # EU559503; from Indonesia), Austrochtonia sp JM 2008 (Genbank accession # EU559513; from Australia) and GenBank voucher sequences of Hyiidae (one specimen) and Olpiidae (two specimens) were included in the latter analysis as follows: Indohya sp JM 2008 (Genbank accession # EU559564) and Beierolpium bornemisszai (Genbank accession #EU559545) and Calocheiridius termitophilus (GenBank accession # EU559544). In addition, as local references, a subset of 46 lineages of Chthoniidae representing 56 specimens, 20 specimens of Hyiidae and 11 specimens of Olpiidae were included in the analyses respectively. Sequences of the scorpion Pandinus imperator (Genbank accession # AY1565821) and harvestman spider Siro rubens (Genbank accession # DQ5131111) were used as outgroups.

Phylogenetic analyses Two phylogenetic analyses were conducted, one for the Chthoniidae and one combined for the Hyiidae/Olpiidae. The phylogenetic analysis of the Chthoniidae, which included the five Chthoniidae lineages from Robe Valley, in addition to 55 reference specimens, placed the

3 Molecular systematics of subfauna from the Robe Valley

Robe Valley specimens in five distinct genetic lineages (Figure 3). The Robe Valley Chthoniidae lineages were not placed in lineages containing reference specimens and therefore were assigned to the new lineages PC047 – PC051. However the lineages showed significant although distant relationships with some reference specimens (Figure 3). Specifically, lineages PC048 and PC051 formed a well-supported clade with one another and with specimen IV203 from deposit 2403A from a previous survey in the Robe Valley (Figure 3). Similarly, lineages PC049 and PC050 formed a well-supported clade with specimen IV210 from Highway/Tod bore from a previous survey at Robe Valley (Figure 3). Finally, lineage PC047 formed a well-supported clade with specimen IS10 from Red Hill (Figure 3).

The phylogenetic analysis of the Hyiidae and Olpiidae, which included the nine lineages from Robe Valley, in addition to 21 reference specimens of Hyiidae and 12 reference specimens of Olpiidae, placed the Robe Valley specimens in nine distinct genetic lineages (Figure 4). Seven lineages were placed in a clade containing the Pilbara and Genbank reference specimens of Hyiidae (Figure 4). The Robe Valley lineages of Hyiidae were not placed in lineages containing reference specimens, therefore were assigned to the new lineages PH019 – PH025 (Figure 4). The Robe Valley Hyiidae lineages did however show significant although distant relationships to some reference specimens (Figure 4). Specifically, lineage PH019 formed a well-supported clade with specimen IV202 from a previous survey at the Robe Valley and lineages PH020 – PH025 formed a well-supported clade with one another and with specimen IV219 from a previous survey at Robe Valley (Figure 4).

Two lineages were placed in a clade containing the Pilbara and Genbank reference specimens of Olpiidae (Figure 4). The Robe Valley Olpiidae lineages were not placed in lineages containing reference specimens, therefore were assigned to the new lineages PO012 and PO013 (Figure 4). The Robe Valley Olpiidae lineages did however show significant although distant relationships to some reference specimens (Figure 4). Specifically, lineage PO012 formed a well-supported clade with specimens IS03 and JF59 from Red Hill and Mesa H, respectively and lineage PO013 formed a well-supported clade with specimens from Munjina, Marillana Creek and Yeelirrie (Figure 4).

Differentiation within and between lineages The fourteen lineages of Pseudoscorpiones differed from one another by a mean of 3.6 to 35.3% sequence divergence (Table 4). Variation within lineages ranged from between a mean of 0.0 and 1.8% sequence divergence (Table 5).

The five lineages of Chthoniidae from the Robe Valley differed from one another by between a mean of 10.4 and 21.2% sequence divergence and from the reference lineages by between 8.5 and 27.1% sequence divergence (Table 6).

The seven lineages of Hyiidae from the Robe Valley differed from one another by between a mean of 3.5 and 21.6% sequence divergence, and from the reference specimens of Hyiidae by between 4.2 and 34.9 % sequence divergence (Table 7).

The two lineages of Olpiidae from the Robe Valley differed from one another by 24.2% sequence divergence and from the reference specimens of Olpiidae by between 14.4 and 27.0% sequence divergence (Table 8).

Schizomids Preliminary analysis - Reference sequences and outgroups Eighteen schizomids (phase 3 and historical) were sequenced from the Robe River Valley (Table 9). In addition, two sequences of Bamazomus subsolanus were provided by the WAM in order to aid identifications. In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted with the 16 Robe River Valley specimens for which sequences were obtained, as well as the B. subsolanus sequences to identify the number of genetic lineages present. A representative from each lineage was analysed with >500 reference sequences of schizomids from previous surveys in the Pilbara to compile a manageable and relevant reference data set. The reference sequences were

4 Molecular systematics of subfauna from the Robe Valley

selected based on the criteria that they showed ≤ 15% sequence divergence from at least one of the Robe River Valley lineages.

Five distinct genetic lineages were identified (Figure 5) and a representative from each was analysed in a model-based phylogenetic analysis with 37 reference specimens. A specimen of schizomid from the family Hubbardiidae, Brignolozomus woodwardi (GenBank accession # EU272675) was used as an outgroup.

Phylogenetic analysis The phylogenetic analysis, which included the five Robe River Valley lineages, in addition to 37 reference specimens of schizomids, placed the Robe River Valley specimens in five distinct lineages, three of which also contained reference specimens from previous surveys at the Robe River Valley (Figure 6). These three were assigned to the existing lineages Paradraculoides anachroretus (SCH036), P. bythius (SCH001) and Robe Valley Bamazomus lineage SCH005; Figure 6). A fourth Robe Valley lineage formed a close relationship with Paradraculoides anachoretus, forming a well-supported clade with that species and was assigned to the new lineage SCH034 (Figure 6). The remaining lineage (Bamazomus subsolanus) formed a well- supported clade with Robe Valley lineage SCH005 from a previous survey (Figure 6).

Differentiation within and between lineages The five lineages of schizomids detected at the Robe River Valley differed from one another by between 4.8 and 19.4% mean sequence divergence (Table 10). Individuals within each of the five lineages differed from one another by between 0.1 and 0.4% mean sequence divergence (Table 11).

Three of the five lineages differed from the closest reference lineages (P. bythius, P. anachoretus, and Robe Valley lineage SCH005), collected during previous surveys at the Robe River Valley, by <1% sequence divergence (Table 12). Lineage SCH034 differed from the closest reference lineage (P. anachoretus) by 6.3% (Table 12) and averaged 4.8% sequence divergence from lineage SCH036 (Table 12). The lineage of B. subsolanus differed from the nearest reference specimens by >15% (Table 12).

Thysanura Reference specimens and outgroups Fourteen of nineteen specimens of Thysanura (historical, phase 3 and phases 1 and 2 repeats) were successfully sequenced for 12s (Table 13). A search of similar sequences on Genbank and in the Helix database indicated that two of the specimens most closely matched database specimens of the subfamily Atelurinae, five of the specimens most closely matched database specimens of Nicoletiinae and one most closely matched specimens of the subfamily Subnicoletiinae. The remaining six specimens did not show close relationships with any database specimens of Thysanura, instead showing the closest (although distant) matches to Plecoptera. Based on those results, GenBank voucher sequences were included as reference sequences of Atelurinae (one specimen) and Nicoletiinae (two specimens) as follows: Atelura formicaria (Genbank accession # EU084035) and Trinemura callawa and T. cundalin (GenBank accession #JQ282164 and JQ282165). In addition, as local references, 11 lineages of Ateluriinae, representing 36 specimens, 16 lineages of Nicoletiinae representing 38 specimens and four lineages of Subnicoletiinae, representing seven specimens were included in the analysis. Two specimens of Hemiptera were used as outgroups: Palomena prasina (Genbank accession # JQ029134) and Graphosoma lineatum (Genbank accession # KM013315)

Phylogenetic analyses The phylogenetic analysis, which included the 14 new specimens from Robe Valley, in addition to 32 reference lineages, placed the Robe Valley specimens in eight distinct genetic lineages (Figure 7).

One of those lineages, containing two specimens, was placed in the clade containing reference lineages of Ateluriinae and was assigned to the new lineage TA009 as it was distinct from the reference lineages (Figure 7). However, Ateluriinae lineage TA009 formed a well- supported clade with a lineage of Ateluriinae from the Cape Preston area (Figure 7).

5 Molecular systematics of subfauna from the Robe Valley

Three of the lineages, containing five specimens in total, were placed in the clade containing reference lineages of Nicoletiinae and were assigned to the new lineages TN010, TN011, TN012, as they were distinct from the reference lineages (Figure 7). All three lineages were placed in a well-supported clade containing reference lineages from Christmas Creek, Barrow Island, and Wheelarra Hill, some of which were assigned to the genus Trinemura on the basis of morphology (Figure 7).

One lineage, containing a single specimen, was placed in the clade containing reference lineages of Hemitrinemura, from the subfamily Subnicoletiinae (Figure 7). It formed a well- supported clade with specimens from Western Hub/Christmas Creek, but was assigned to the new lineage TS005, as it was distinct from the reference lineages (Figure 7).

Three lineages, containing six specimens in total, were distinct from the reference lineages of Nicoletiinae, Subnicoletiinae and Ateluriinae, and did not show close matches to any GenBank specimens of Thysanura. These three lineages were assigned to the new lineages T001, T002 and T003; Figure 7).

Differentiation within and between lineages The lineage of Ateluriinae TA009 differed from the nearest reference lineage TA007 by 9.3 to 13.2% sequence divergence (Table 14). The Three lineages of Nicoletiinae differed from one another by 11.1 to 12.9% sequence divergence (Table 15). They differed from the reference lineages by 8.7 to 22.3% sequence divergence (Table 15). The lineage of Subnicoletiinae, TS005 differed from the nearest reference lineage, TS004 by 5.3% and from the remaining reference lineages of Subnicoletiinae by 8.7 to 14.5% sequence divergence (Table 16). The three lineages of the unknown family, T001 – T003 differed from one another by between 3.1 and 19.2% sequence divergence (Table 17).

Distribution of lineages The Ateluriinae lineage TA009 was detected in a single drillhole at Mesa B. Nicoletiinae lineages TN010, TN011 and TN012 were detected at Highway-Tod and Mesa B, deposit 240E, and Mesa C, respectively. The Subnicoletiinae lineage TS005 was detected at Highway-Tod. Lineage T001 of the unknown family was detected at Highway-Tod/Waramboo, and lineages T002 and T003 were detected at Mesa B.

Conclusions COI is widely considered to show suitable variation to distinguish species (Hebert et al., 2003a). In a comparison of COI sequences for over 13,000 pairs of taxa, Hebert et al (2003b) found a mean of 11.1% sequence divergence between distinct species. Nearly 80% of the comparisons showed that species pairs differed from one another by greater than 8% sequence divergence. However, a taxon by taxon approach, examining the amount of phylogenetic variation within and between species is the most widely accepted method of defining species.

Araneae Four genetically distinct lineages of Araneae were detected, likely representing four distinct species, owing to the high level of sequence divergence among them and between them and the reference lineages. The four lineages differed from the reference lineages by >10%, indicating that all are likely to be new lineages and species, based on the material available for comparison. Some of the species may be tentatively assigned to families, however, based on their phylogenetic relationships with reference specimens. Two species may belong to the Oonopidae and one to the Aranidae. The fourth species may belong to the Symphytognathidae, but was placed in a larger clade containing Genbank reference specimens of Theriidae.

Pseudoscorpiones

Fourteen distinct genetic lineages of pseudoscorpions were detected at Robe Valley: five Chthoniidae, seven Hyiidae and two Olpiidae.

6 Molecular systematics of subfauna from the Robe Valley

The five lineages of Chthoniidae differed from one another by >10% sequence divergence, thus each is likely to represent a distinct species. Further, the five Chthoniidae lineages from Robe Valley differed from the reference specimens by >8.5%, indicating that the five are likely to be new species that have so far not been detected in the Pilbara, based on the material available for comparison.

The two lineages of Olpiidae differed from one another by 24.2% sequence divergence, thus each represents a distinct species. Further, the two Olpiidae lineages from Robe Valley differed from the reference specimens by >14%, indicating that the five are likely to be new species that have so far not been detected in the Pilbara, based on the material available for comparison.

Six of the seven lineages of Hyiidae (PH020 – PH025) were closely related, forming a well- supported clade and differing from one another by <10% sequence divergence (range= 3.5 – 9.5%). While these values are relatively low, they are higher than is typically observed between members of the same species. The observed genetic differentiation among these lineages may reflect multiple species, incipient speciation or differentiation within one or a few species, owing to the poor dispersal abilities of this group. Sampling of intermediate sites may help resolve the relationship between these lineages. In contrast, lineage PH019 differed from the remaining Robe Valley and reference lineages by >16% sequence divergence, indicating that it is likely to represent a new species that has not been detected previously in the Pilbara, based on the material available for comparison.

Schizomida Previous analyses of genetic variation between morphologically distinct species of Schizomida can be used as a genetic ‘yardstick’ to interpret the current data set. The five described species of Paradraculoides (Harvey et al, 2008) differ from one another by between 8.4 to 12.1% sequence divergence (uncorrected p-distances; calculated by us from the Harvey et al., 2008 data). Similarly, the four described species of Draculoides differ from one another by between 4.5 to 13.7% sequence divergence (uncorrected p-distances calculated by us from Harvey et al., 2008).

Five genetically distinct lineages of schizomids were detected at the Robe River Valley. The five lineages likely correspond to four or five species. Three were assigned to the existing lineages and species Paradraculoides anachroretus, P. bythius and Robe Valley Bamazomus lineage SCH005, differing from these reference lineages by <1%. A fourth Robe Valley lineage formed a close relationship with Paradraculoides anachoretus, differing from the P. anachoretus lineage by an average of 4.8% sequence divergence. Thus the relationship between these two lineages requires further investigation, as the observed divergence, while moderately low, is greater than that generally observed between individuals of the same species. Sampling from intermediate sites may help to resolve the relationship between these two lineages. The remaining lineage (Bamazomus subsolanus) differed from the nearest reference specimens by >15% and thus is likely to be a new species, which has not been detected in the Pilbara so far, based on the material available for comparison.

Thysanura The mitochondrial gene 12S is widely used in insect systematics (Simon et al., 1996; Caterino et al., 2000), although in contrast to the mitochondrial gene COI, fewer broadscale comparative studies are available to provide a basis for species discrimination. Nonetheless, 12S has proven useful for establishing phylogenetic relationships in many insect groups (Caterino et al., 2000). The 12S gene evolves approximately 1.5 times (Mueller, 2006) more slowly than COI. Hence we would expect that the threshold for species discrimination using 12S would be, by inference, lower than COI.

Eight distinct lineages of Thysanura were detected in the present study, one Ateluriinae (TA009), three Nicoletiinae (TN010, TN011 and TN012), one Subnicoletiinae (TS005) and three belonging to an undetermined family (T001, T002, T003). The eight lineages likely represent seven species, with each lineage corresponding to a species with the exception of lineages T002 and T003 from the unknown family, which differed from one another by an average of just over 3%. Using a conversion factor of 1.5 x to account for the slower 12s rate of evolution, this value might correspond to approximately 4.5% of COI divergence. Given the relatively low value, but

7 Molecular systematics of subfauna from the Robe Valley

keeping in mind that these are estimates based on current understanding of 12s and COI evolution rates, it is likely that lineages T002 and T003 comprise a single species, showing some genetic divergence among drillholes. The taxonomic placement of lineages T001 – T003 is uncertain, owing to the poor match of these sequences to the Helix and Genbank databases.

Six of the seven species are likely to be new as they differed from the nearest reference lineages by approximately > 9% sequence divergence (= approximately 13.5% COI divergence, using the 1.5 conversion factor). Subnicoletiinae lineage TS005 requires further examination. It differed from a reference lineage from Western Hub by approximately 5% sequence divergence at 12s. Again, using a conversion factor of 1.5, this may correspond to approximately 7.5% COI sequence divergence. While this level of differentiation exceeds typical levels of intraspecific variation, it is clear that the lineage is closely related to the lineage from Western Hub. Sampling of intermediate sites may help to determine the relationship between the two lineages.

Summary So far, 31 lineages from four taxonomic groups were detected during the present study at Robe Valley, as shown below. The 31 lineages likely represent 24 - 30 species. Five of the species appear to have been detected previously in the Pilbara, although further investigation may be required to fully resolve relationships. The remaining species at Robe Valley are likely to be new, based on the material available for comparison.

The failure rate was somewhat higher than usual with this project. In addition to our inability to produce amplification products from some historical specimens, which showed evidence of degraded DNA, of those that did amplify, some sequences were contaminated with foreign DNA. This happens when there isn't enough target DNA to out-amplify contaminants.

Araneae Four lineages were detected at Robe Valley, likely corresponding to four species. None of the species have been detected previously in the Pilbara, based on the material available for comparison.

Oonopidae: AO014, AO015 = two species Aranidae: AA001 = one species Symphytognathidae/Theriidae: A012 = one species

Pseudoscorpiones Fourteen lineages were detected at Robe Valley, likely corresponding to 9 - 14 species. None of the species of Chthoniidae and Olpiidae have been detected previously in the Pilbara, based on the material available for comparison. One species of Hyiidae is likely to be new, however, the remaining six Hyiidae lineages form a group that includes a lineage detected previously at Mesa B, and the relationships between these closely related lineages need further investigation

Chthoniidae: PC047, PC048, PC049, PC050, PC051 = five species Olpiidae: PO012, PO013 = two species Hyiidae: PH019 = one species Hyiidae: PH020, PH021, PH022, PH023, PH024, PH025 = one to six species

Schizomida Five lineages were detected at Robe Valley, likely corresponding to four or five species. Three of the lineages were placed with reference specimens of Paradraculoides, and two were placed with reference specimens of Bamazomus. Three of the species have been detected previously in the Pilbara (lineage SCH005 (Bamazomus), P. bythius and P. anachoretus). A fourth shows moderate genetic similarity to P. anachoretus and requires further investigation, while the remaining species of Bamazomus appears to be new, based on the material available for comparison.

Paradraculoides: SCH001 (P. b ythius), SCH036 (P. anachoretus), SCH034 (similar to P. anachoretus)= two or three species

8 Molecular systematics of subfauna from the Robe Valley

Bamazomus: SCH005, SCH035 = two species

Thysanura Eight lineages were detected at Robe Valley, likely corresponding to seven species in four subfamilies. Six of the species appear to be new, owing to the large distances between them and the reference specimens available for comparison. However one species, TS005, appears to be closely related to a species that was detected previously at Western Hub.

Ateluriinae: TA009 = one species Nicoletiinae: TN010, TN011 and TN012 = three species Subnicoletiinae TS005 = one species undetermined family T001, T002, T003 = two species are likely, T001 and T002+T003

References

9 Molecular systematics of subfauna from the Robe Valley

Table 1. Specimens of Araneae used in the present study and the lineage to which they were assigned, based on variation at the COI gene. nd=no data. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat. Specimen_ID Drillhole_ID Provenance Cap No. Helix ID Lineage SSp-2010-253 DCBRC-017 H IV345 AO014 SSp-2010-75 DCBRC-040 H IV346 AO014 MEBRC0016P1T1-2 MEBRC0016 H 155 IV272 AO015 110817 H IV273 contam 127038 H IV274 No amp 127039 H IV275 AA001 138441 H IV276 A012

Table 2 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Araneae detected at Robe Valley and the reference lineages as shown in Figure 1. Distances between the Robe Valley specimens and the reference specimens are highlighted in yellow.

Table 3. Specimens of Pseudoscorpiones used in the present study and the lineage to which they were assigned, based on variation at the COI gene. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat.

Specimen ID Drillhole ID Provenance Helix ID Lineage 99264 H IV277 PO012 127104 H IV278 PC051 127105 H IV279 PC048 127106 H IV280 PC048 132672 H IV281 PO013 132684 H IV282 contam WAM 127103 K09960712T1-4 H aliq PH019 DD15MEB0018-20160317-T2-01 DD15MEB0018 P3 IV258 messy DD15MEB0018-20160317-T3-01 DD15MEB0018 P3 IV259 PHO023 GR15MEB0014-20160317-T1-01 GR15MEB0014 P3 IV260 messy GR15MEB0022-20160115-sc-01 GR15MEB0022 P3 IV261 PC047 GR15MEB0022-20160115-sc-02 GR15MEB0022 P3 IV262 PC047 GR15MEB0022-20160115-sc-03 GR15MEB0022 P3 IV263 PC047 GR15MEB0022-20160115-sc-04 GR15MEB0022 P3 IV264 PC047 RC15MEB0020-20160317-T2-02 RC15MEB0020 P3 IV265 PHO021 RC15MEB0020-20160317-T3-01 RC15MEB0020 P3 IV266 PHO021 RC15MEB0171-20160317-T2-01 RC15MEB0171 P3 IV267 messy RC15MEB0171-20160317-T2-02 RC15MEB0171 P3 IV268 PC049 RC15MEB0171-20160317-T3-01 RC15MEB0171 P3 IV269 PC050 RC15MEB0171-20160317-T3-02 RC15MEB0171 P3 IV270 PC049 138532 RC14MEB0010 Re IV216 PHO024 138536 RC14MEB0115 Re IV220 NoAmp 138537 RC14MEB0123 Re IV222 PHO025 138538 RC14MEB0123 Re IV224 PHO022 139896 RC14MEB0115 Re IV221 PHO020 139897 RC14MEB0123 Re IV223 PHO025 139898 RC14MEB0123 Re IV225 PHO025

10 Molecular systematics of subfauna from the Robe Valley

Table 4. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Pseudoscorpiones detected in the NJ analysis as shown in Figure 2. Distances between lineages within each family are highlighted in yellow.

Chthoniidae Olpiidae Hyiidae Lineage 1 2 3 4 5 6 7 8 9 10 11 12 13 14 1 0.012 0.014 0.014 0.016 0.016 0.015 0.016 0.019 0.017 0.019 0.017 0.016 0.016 2 0.117 0.013 0.014 0.014 0.016 0.017 0.017 0.021 0.018 0.019 0.017 0.016 0.016 3 0.165 0.154 0.011 0.014 0.016 0.016 0.016 0.020 0.017 0.018 0.016 0.014 0.015 4 0.174 0.174 0.104 0.013 0.017 0.016 0.015 0.019 0.016 0.018 0.016 0.015 0.016 5 0.213 0.186 0.194 0.178 0.015 0.018 0.017 0.020 0.016 0.018 0.017 0.016 0.016 6 0.278 0.253 0.263 0.269 0.261 0.015 0.013 0.019 0.014 0.017 0.014 0.014 0.013 7 0.239 0.235 0.251 0.256 0.249 0.242 0.017 0.020 0.017 0.018 0.016 0.016 0.016 8 0.279 0.263 0.294 0.278 0.249 0.297 0.263 0.019 0.016 0.016 0.015 0.014 0.014 9 0.332 0.329 0.353 0.342 0.326 0.334 0.316 0.249 0.015 0.015 0.015 0.015 0.013 10 0.274 0.248 0.276 0.275 0.261 0.272 0.234 0.206 0.153 0.010 0.009 0.011 0.008 11 0.270 0.240 0.280 0.279 0.249 0.286 0.220 0.214 0.164 0.078 0.007 0.011 0.008 12 0.273 0.254 0.276 0.277 0.265 0.278 0.241 0.206 0.154 0.070 0.036 0.010 0.006 13 0.279 0.250 0.279 0.264 0.256 0.267 0.247 0.216 0.168 0.095 0.075 0.069 0.007 14 0.269 0.252 0.275 0.267 0.254 0.264 0.242 0.201 0.150 0.069 0.047 0.035 0.039

11 Molecular systematics of subfauna from the Robe Valley

Table 5. Mean distance (D) and standard error (s.e.) within lineages of Pseudoscorpiones detected in the NJ analysis as shown in Figure 2 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis.

Lineage D s.e. N rep 1 0.000 0.000 2 IV268 2 n/c n/c 1 IV269 3 n/c n/c 1 IV278 4 0.001 0.001 2 IV279 5 0.001 0.001 4 IV261 6 n/c n/c 1 IV277 7 n/c n/c 1 IV281 8 n/c n/c 1 127103 9 n/c n/c 1 IV221 10 0.001 0.001 2 IV265 11 n/c n/c 1 IV224 12 n/c n/c 1 IV259 13 n/c n/c 1 IV216 14 0.018 0.004 3 IV222

Table 6 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Chthoniidae (Pseudoscorpiones) detected at Robe Valley and the reference lineages as shown in Figure 3. Distances between the Robe Valley specimens and the reference specimen are highlighted in yellow.

Table 7 (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Hyiidae (Pseudoscorpiones) detected at Robe Valley and the reference lineages as shown in Figure 4. Distances between the Robe Valley specimens and the reference specimen are highlighted in yellow.

12 Molecular systematics of subfauna from the Robe Valley

Table 8. Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Olpiidae (Pseudoscorpiones) detected at Robe Valley and the reference lineages as shown in Figure 4. Distances between the Robe Valley specimens and the reference specimens are highlighted in yellow.

Lineage IV277 IV281 BQ20 Cter Bbor G320 G322 G323 G325 G326 IS03 JD02 JF59 G89 G90 IV277 99264 Pseudoscorpiones 0.016 0.019 0.019 0.021 0.019 0.020 0.020 0.020 0.018 0.014 0.020 0.018 0.018 0.019 IV281 132672 Pseudoscorpiones 0.242 0.021 0.019 0.020 0.017 0.017 0.018 0.016 0.017 0.020 0.020 0.019 0.019 0.020 BQ20 PES 1896 Beierolpium won1 0.254 0.272 0.021 0.019 0.022 0.018 0.017 0.019 0.021 0.021 0.022 0.016 0.020 0.022 EU559544 Calocheiridius termitophilus 0.235 0.221 0.242 0.018 0.019 0.020 0.019 0.019 0.020 0.020 0.020 0.021 0.020 0.019 EU559545 Beierolpium bornemisszai 0.253 0.241 0.230 0.194 0.020 0.019 0.021 0.019 0.018 0.019 0.020 0.017 0.020 0.019 G320 LN8818 Olpiidae sp MJ 0.263 0.176 0.308 0.232 0.246 0.017 0.015 0.015 0.016 0.021 0.019 0.019 0.016 0.017 G322 100795 Olpiidae sp MJ 0.270 0.144 0.263 0.220 0.229 0.137 0.017 0.016 0.016 0.018 0.017 0.017 0.019 0.017 G323 100292 Olpiidae sp MJ 0.249 0.181 0.269 0.235 0.246 0.176 0.160 0.016 0.013 0.019 0.021 0.019 0.020 0.018 G325 LN9467 Olpiidae sp MA 0.252 0.176 0.265 0.211 0.220 0.164 0.144 0.128 0.016 0.019 0.019 0.019 0.019 0.017 G326 LN7150 Olpiidae sp MC 0.270 0.174 0.285 0.227 0.229 0.139 0.139 0.148 0.148 0.019 0.018 0.017 0.018 0.016 IS03 Nolabel-20150709-T1-02 0.205 0.274 0.253 0.224 0.228 0.251 0.242 0.281 0.267 0.247 0.017 0.017 0.019 0.018 JD02 RC12KOOD2579-151010-T4-02 0.247 0.241 0.269 0.200 0.193 0.215 0.201 0.215 0.219 0.228 0.233 0.020 0.018 0.019 JF59 RC15MEH0302-20151210-T1-03 0.174 0.247 0.244 0.221 0.211 0.247 0.240 0.226 0.244 0.260 0.220 0.233 0.018 0.019 YYAC26 G89 LN7692 0.255 0.183 0.269 0.198 0.258 0.164 0.171 0.192 0.173 0.180 0.269 0.241 0.251 0.018 YYHC0048L G90 LN7592 0.252 0.178 0.315 0.234 0.264 0.165 0.156 0.182 0.180 0.158 0.269 0.241 0.271 0.169

13 Molecular systematics of subfauna from the Robe Valley

Table 9. Specimens of Schizomida used in the present study and the lineage to which they were assigned, based on variation at the COI gene. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat.

Cap Specimen ID Drillhole ID Provenance No. Helix ID Lineage 65760 H IV283 No amp 65761 H IV284 P. bythius 65762 H IV285 Messy seq 92222 H 182 IV286 P. bythius 133444 H IV287 P. bythius 110826 H IV288 P. bythius 110827 H IV289 P. bythius 133445 H IV290 P. bythius 110830 H IV291 P. bythius 140989 H IV292 P. bythius 140982 H IV293 SCH034 140983 H IV294 SCH034 P. 133446 H IV295 anachoretus B. 137066 WAM seq 137066 subsolanus B. 137067 WAM seq 137067 subsolanus DD15MEB0008-20160317-T2-01 DD15MEB0008 P3 165 IV326 SCH005 DD15MEB0018-20160317-T3-02 DD15MEB0018 P3 73 IV327 P. bythius GR15MEB0004-20160317-T1-01 GR15MEB0004 P3 170 IV328 P. bythius RC14MEB0102-20160317-T3-06 RC14MEB0102 P3 46 IV329 P. bythius RC15MEB0020-20160317-T3-02 RC15MEB0020 P3 220 IV330 P. bythius

Table 10. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Schizomida detected in the NJ analysis as shown in Figure 5.

Lineage 1 2 3 4 5 1 0.012 0.010 0.014 0.015 2 0.100 0.007 0.012 0.014 3 0.083 0.048 0.012 0.015 4 0.157 0.152 0.145 0.015 5 0.194 0.189 0.185 0.171

Table 11. Mean distance (D) and standard error (s.e.) within lineages of Schizomida detected in the NJ analysis as shown in Figure 5 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis.

Lineage D s.e. N rep 1 0.004 0.002 12 IV289 2 n/c n/c 1 IV295 3 0.001 0.001 2 IV293

14 Molecular systematics of subfauna from the Robe Valley

4 n/c n/c 1 IV326 5 0.002 0.002 2 WAMT137066

Table 12. (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Schizomida detected at Robe Valley and the reference lineages as shown in Figure 6. Distances between the Robe Valley specimens and the reference specimen are highlighted in yellow.

Table 13. Specimens of Thysanura used in the present study and the lineage to which they were assigned, based on variation at the COI gene. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat. Helix Lineage Specimen_ID Drillhole_ID Provenance Cap No. ID MEGRC130P1T1-1 MEGRC130 H 66 IV331 T003 A1778T1-2 A1778 H IV332 No amp MEBRC0020T1-20 MEBRC0020 H IV333 TN010 M2ERC0057T1-21 M2ERC0057 H IV334 TN011 MEAD2517AT1-22 MEAD2517A H IV335 No amp BC186-204023-02 BC186 H 30 IV336 No amp TOBRC0055P1T1-1 TOBRC0055 H 379 IV337 T001 GR15MEC0001-20160316-T2-06 GR15MEC0001 P3 59 IV338 TN012 RC15MEC0197-20160316-T2-01 RC15MEC0197 P3 90 IV339 TN012 GR15MEB0003-20150115-SC-01 GR15MEB0003 P3 5 IV340 T003 GR15MEB0009-20150115-SC-01 GR15MEB0009 P3 6 IV341 T003 GR15MEB0014-Sc-02 GR15MEB0014 P3 10 IV342 T003 GR15MEC008-20160116-sc-01 GR15MEC0008 P3 8 IV343 No amp RC15MEB0171-20150115-SC-01 RC15MEB0171 P3 7 IV344 T002 IV227 MEARC4383 P1/2 re IV227 TN010 IV228 RC14MEB0060 P1/2 re IV228 TA009 IV229 RC14MEB0060 P1/2 re IV229 TA009 IV230 MEARC3814 P1/2 re IV230 TS005

15 Molecular systematics of subfauna from the Robe Valley

Table 14. Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Ateluriinae (Thysanura) detected at Robe Valley and the reference lineages as shown in Figure 7. Distances between the Robe Valley specimens and the reference specimens are highlighted in yellow.

Lineage IV228 IV229 Aform EA0118R BQ14 GA0011R EXR1139 G313 BX3 A32 NB1 BQ13 EEX0078 WRKRC148 IV228 RC14MEB0060 TA009 0.000 0.025 0.018 0.019 0.018 0.019 0.019 0.019 0.015 0.014 0.019 0.017 0.018 IV229 RC14MEB0060 TA009 0.000 0.025 0.020 0.020 0.019 0.020 0.020 0.020 0.016 0.016 0.019 0.019 0.019 EU084035 Atelura formicaria 0.307 0.320 0.025 0.026 0.022 0.024 0.024 0.025 0.024 0.022 0.024 0.023 0.023 EA0118R OB24 TA001 0.157 0.173 0.249 0.011 0.011 0.013 0.014 0.020 0.019 0.018 0.010 0.011 0.010 BQ14 PES 5086 Atelurinae won1 TA002 0.163 0.175 0.314 0.055 0.014 0.014 0.012 0.020 0.017 0.018 0.013 0.014 0.014 GA0011R Jirrpalpur Range TA003 0.158 0.176 0.241 0.052 0.067 0.013 0.015 0.021 0.018 0.017 0.012 0.013 0.011 EXR1139 Alligator Range TA004 0.154 0.170 0.256 0.078 0.093 0.073 0.014 0.019 0.019 0.017 0.013 0.014 0.015 G313 101035 Ateluridae MA TA005 0.173 0.184 0.295 0.078 0.076 0.096 0.064 0.018 0.019 0.017 0.013 0.013 0.013 BX3 PES-5124 Atelurinae MH1 TA006 0.169 0.185 0.312 0.181 0.176 0.184 0.169 0.159 0.023 0.022 0.020 0.020 0.020 A32 TA007 0.114 0.132 0.285 0.171 0.183 0.166 0.169 0.180 0.218 0.009 0.020 0.018 0.019 NB1 TA007 0.093 0.112 0.259 0.170 0.170 0.152 0.155 0.152 0.207 0.053 0.019 0.016 0.016 BQ13 PES 4627 Atelurinae won1 TA008 0.166 0.180 0.302 0.038 0.078 0.067 0.087 0.086 0.162 0.166 0.161 0.012 0.011 EEX0078 Jinaryi TA010 0.145 0.158 0.247 0.052 0.073 0.070 0.078 0.073 0.163 0.151 0.135 0.055 0.008 WRKRC148 J7-7 TA011 0.151 0.164 0.246 0.049 0.070 0.061 0.093 0.087 0.172 0.153 0.137 0.047 0.032

16 Molecular systematics of subfauna from the Robe Valley

Table 15. Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Nicoletiinae (Thysanura) detected at Robe Valley and the reference lineages as shown in Figure 7. Distances between the Robe Valley specimens and the reference specimens are highlighted in yellow.

Lineage IV227 IV333 IV334 IV338 IV339 Tcall Tcund G318 PROP003 G126 G206 AY2 BX42 BX43 CQ10 CQ11 EJ0245R CA35 FL3 EXR0983 CQ13

IV227 MEARC4383 TN010 0.007 0.018 0.018 0.018 0.017 0.018 0.019 0.015 0.017 0.015 0.022 0.015 0.016 0.016 0.020 0.020 0.015 0.014 0.015 0.018

IV333 MEBRC0020T1-20 TN010 0.028 0.017 0.018 0.017 0.019 0.019 0.019 0.016 0.016 0.014 0.023 0.015 0.015 0.016 0.019 0.019 0.015 0.015 0.013 0.017

IV334 M2ERC0057T1-21 TN011 0.129 0.124 0.015 0.015 0.020 0.020 0.018 0.021 0.019 0.017 0.024 0.017 0.015 0.018 0.021 0.021 0.017 0.017 0.020 0.019

IV338 GR15MEC0001 TN012 0.116 0.119 0.124 0.006 0.021 0.019 0.017 0.018 0.018 0.018 0.022 0.017 0.016 0.016 0.018 0.018 0.016 0.015 0.021 0.019

IV339 RC15MEC0197 TN012 0.111 0.113 0.121 0.017 0.020 0.018 0.016 0.018 0.018 0.018 0.022 0.017 0.015 0.016 0.018 0.018 0.015 0.014 0.020 0.019

JQ282164 Trinemura callawa 0.154 0.151 0.187 0.159 0.159 0.013 0.018 0.021 0.019 0.019 0.021 0.018 0.019 0.021 0.019 0.021 0.019 0.020 0.020 0.020

JQ282165 Trinemura cundalin 0.169 0.166 0.202 0.162 0.162 0.065 0.018 0.022 0.020 0.020 0.021 0.019 0.019 0.021 0.018 0.021 0.019 0.019 0.018 0.020

G318 LN8588 MC TN001 0.207 0.190 0.195 0.196 0.190 0.135 0.144 0.021 0.017 0.018 0.018 0.018 0.019 0.020 0.019 0.021 0.016 0.017 0.016 0.020

PROP003 J16-2 TN002 0.155 0.152 0.174 0.188 0.182 0.196 0.193 0.176 0.019 0.020 0.023 0.018 0.018 0.021 0.021 0.022 0.019 0.017 0.019 0.018

G126 TN003 0.175 0.172 0.169 0.196 0.193 0.193 0.190 0.164 0.140 0.016 0.025 0.018 0.016 0.018 0.017 0.019 0.018 0.017 0.019 0.016

G206 108042 TN004 0.141 0.134 0.171 0.172 0.171 0.182 0.182 0.196 0.159 0.099 0.022 0.013 0.014 0.019 0.018 0.019 0.016 0.015 0.017 0.016

AY2 BES16311 B10 TN005 0.198 0.194 0.215 0.190 0.193 0.173 0.187 0.115 0.197 0.231 0.209 0.023 0.023 0.023 0.020 0.023 0.022 0.020 0.021 0.021

BX42 PES-5266 MH1 TN006 0.134 0.120 0.134 0.162 0.156 0.156 0.165 0.204 0.167 0.172 0.164 0.229 0.014 0.019 0.018 0.017 0.016 0.016 0.017 0.018

BX43 PES-5964 MH1 TN007 0.169 0.144 0.153 0.169 0.163 0.170 0.167 0.176 0.131 0.129 0.126 0.182 0.137 0.016 0.018 0.018 0.014 0.015 0.017 0.018

CQ10 B18 CC1886 TN008 0.104 0.124 0.132 0.101 0.098 0.190 0.222 0.229 0.198 0.190 0.174 0.203 0.175 0.192 0.022 0.020 0.015 0.015 0.018 0.019

CQ11 B15 CC0179 TN009 0.181 0.171 0.185 0.200 0.194 0.160 0.156 0.161 0.171 0.121 0.154 0.170 0.171 0.151 0.203 0.019 0.018 0.018 0.020 0.018

EJ0245R J2 36 OB18 TN011 0.188 0.185 0.172 0.199 0.196 0.210 0.183 0.187 0.158 0.148 0.155 0.221 0.190 0.163 0.201 0.128 0.021 0.016 0.018 0.019

CA35 11:1513 TN013 0.121 0.121 0.141 0.114 0.111 0.152 0.167 0.199 0.183 0.194 0.180 0.195 0.153 0.162 0.155 0.189 0.188 0.015 0.018 0.020

FL3 TOBRC0011PIT2-1 A TN014 0.092 0.098 0.120 0.092 0.087 0.168 0.177 0.209 0.170 0.162 0.138 0.217 0.139 0.153 0.101 0.175 0.172 0.109 0.017 0.017

EXR0983 J8 13 Jimblebar W TN015 0.097 0.094 0.128 0.137 0.134 0.180 0.180 0.160 0.181 0.188 0.180 0.204 0.144 0.165 0.124 0.166 0.169 0.128 0.125 0.019

CQ13 B19 CC0748 TN016 0.209 0.193 0.223 0.214 0.209 0.152 0.165 0.188 0.163 0.132 0.167 0.188 0.207 0.190 0.224 0.119 0.146 0.205 0.202 0.176

17 Molecular systematics of subfauna from the Robe Valley

Table 16. Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Subnicoletiinae (Thysanura) detected at Robe Valley and the reference lineages as shown in Figure 7. Distances between the Robe Valley specimens and the reference specimens are highlighted in yellow.

Lineage IV230 KCRC0107 FH4 FH5 GH1 IV230 MEARC3814 TS004 0.017 0.016 0.012 0.012 KCRC0107 TS001 0.145 0.019 0.015 0.015 FH4 Hemitrinemura sp B10 TM0087 TS002 0.089 0.132 0.015 0.016 FH5 Hemitrinemura sp B10 BG0082 TS003 0.087 0.118 0.101 0.012 GH1 Trinemura Tri-PES14085 TS004 0.053 0.124 0.092 0.074

Table 17. Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of the unknown family of Thysanura detected at Robe Valley as shown in Figure 7.

Lineage T001 T002 T003

Specimen IV337 IV344 IV331 IV340 IV341 IV342 IV337 TOBRC0055P1T1-1 T001 0.016 0.019 0.017 0.016 0.015 IV344 RC15MEB0171 T002 0.157 0.010 0.009 0.008 0.008 IV331 MEGRC130P1T1-1 T003 0.192 0.038 0.006 0.006 0.006 IV340 GR15MEB0003 T003 0.178 0.037 0.013 0.000 0.000 IV341 GR15MEB0009 T003 0.168 0.033 0.013 0.000 0.000 IV342 GR15MEB0014-Sc-02 T003 0.160 0.031 0.013 0.000 0.000

18 Molecular systematics of subfauna from the Robe Valley

Figure 1. Bayesian analysis of COI haplotypes of Araneae from the present study and reference specimens from Genbank and previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

KC708097 Missulena sp. WAM T97637 Actinopodidae Mygal KC708079 Missulena sp. WAM T113626 Actinopodidae Mygal IV346 SSp 2010 75 DCBRC 040 Araneae AO014 1.00 0.55 IV345 SSp 2010 253 DCBRC 017 Araneae AO014 DF9 8999 Araneomorphae sp. AO010 G308 LN9827 Oonopidae sp MJ AO006 1.00 G307 LN9925 Gnaphosidae sp MJ AO007 0.81 EO8 Prethopalpus sp EMP0049 AO011 0.97 1.00 EO7 Prethopalpus sp EMP0061 x2 AO012 1.00 EO33 Prethopalpus sp ECP0057 AO013 IV272 MEBRC0016P1T1 2 Araneae AO015 1.00 0.80 KR864743 Ischnothyreus auritus Oonopidae G141 Prethopalpus sp. S2 LN8445 x2 A0004 G140 Oonopidae sp. indet. LN8185 AO001 0.65 1.00 0.93 G135 Prethopalpus sp. S2 LN7377 x2 AO001 0.94 0.90 G136 Oonopidae sp. indet. LN7327 x2 AO001 1.00 G139 Oonopidae sp. indet. LN8432 AO002 G137 Oonopidae sp. indet. 100075 AO003 G309 LN8606 Prethopalpus sp MA AO005 CA6 11 1510 Araneae indet. AO009 IV276 138441 Araneae A012

0.57 G226 Araneae 107786 A008 1.00 0.74 G210 Araneae 107945 A008 0.53 DF8 8662 Anapistula sp. A010 1.00 BX1 PES 5149 Anapistula MH1 Symphytognathidae A009 1.00 G227 Araneae MC8638 A006 0.96 0.62 G209 Araneae 109675 A007 1.00 GU684645 Theridion ohlerti Theridiidae AY231053 Robertus neglectus Theriidae 1.00 IV275 127039 Araneae AA001 1.00 0.76 KJ957946 Arachnura scorpionoides Aranidae 1.00 KJ957966 Gea theridioides Aranidae FG1100 Wheelarra Hill A002 1.00 0.99 1.00 EJ0003R OB18 A002 BP013 Deposit B x2 A001 1.00 GH12 Selinopidae PES16307 A011 G145 Desognanops sp. n. Y1 LN6635 x2 A004 1.00 G142 Desognanops sp. n. Y1 LN8482 A005 0.1

19 Molecular systematics of subfauna from the Robe Valley

Figure 2. Neighbour-joining analysis of specimens of Pseudoscorpiones from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

IV268 RC15MEB0171-20160317-T2-02 Pseudoscorpiones 1 IV270 RC15MEB0171-20160317-T3-02 Pseudoscorpiones IV269 RC15MEB0171-20160317-T3-01 Pseudoscorpiones 2 IV278 127104 Pseudoscopiones 3 IV279 127105 Pseudoscopiones 4 IV280 127106 Pseudoscopiones IV264 GR15MEB0022-20160115-sc-04 Pseudoscorpiones IV261 GR15MEB0022-20160115-sc-01 Pseudoscorpiones 5 IV262 GR15MEB0022-20160115-sc-02 Pseudoscorpiones IV263 GR15MEB0022-20160115-sc-03 Pseudoscorpiones

IV277 99264 Pseudoscopiones 6 IV281 132672 Pseudoscopiones 7 127103 Pseudoscorpiones 8 IV221 139896 Pseudoscorpiones 9 IV265 RC15MEB0020-20160317-T2-02 Pseudoscorpiones 10 IV266 RC15MEB0020-20160317-T3-01 Pseudoscorpiones IV224 138538 Pseudoscorpiones 11 IV259 DD15MEB0018-20160317-T3-01 Pseudoscorpiones 12 IV216 138532 Pseudoscorpiones 13 IV222 138537 Pseudoscorpiones IV223 139897 Pseudoscorpiones 14 IV225 139898 Pseudoscorpiones

0.02

20 Molecular systematics of subfauna from the Robe Valley

Figure 3. Bayesian analysis of COI haplotypes of Chthoniidae (Pseudoscorpiones) from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site. AY156582 Pandinus imperator DQ513111 Siro rubens Anaulacodithella sp. EU559520 0.62 0.68 EU559521 Sathrochthonius insulanus EU559519 Pseudochthonius sp. EU559506 Tyrannochthonius sp. JM 2008 EU559503 Lagynochthonius johni EU559509 Pseudotyrannochthonius sp. JM 2 0.74 G146 Pseudoscorpians LN6643 PC016 0.87 G149 Pseudoscorpians LN8500 PC017 0.60 1.00 0.87 G154 Pseduoscorpians LN8374 PC018 G152 Pseudoscorpians LN7608 PC020 1.00 1.00 SB14 MT G85 LN8520 PC026 G151 Pseudoscorpians LN8451 PC019 1.00 YYHC0048I G87 LN8425 PC034 EU559505 Paraliochthonius sp. JM 2008 KJ659959 Tyrannochthonius aridus IV212 DD14MEL0001 20151001 T2 02 PC009 0.69 1.00 0.99 IV226 RC14MRR0020 20151002 T2 01 PC010 0.64 EAP0178 Tyran B1 Ophthalmia Range PC032 AX83 10DDH033P1T1 2 PC042 0.81 0.60 IS01 PC001 1.00 IS12 PC002 0.99 1.00 1.00 IS08 PC003 0.73 IS06 PC004 1.00 0.99 IS05 PC005 JV08 DD15BS40010 20160608 T2 02 PC046 0.71 MG0165R J8 31 Tyrannochthonius sp PC035 G219 Pseudoscorpian 108935 Turner River PC022 1.00 G220 Pseudoscorpian 109177 Turner River PC023 0.54 0.67 1.00 G479 111446 Chthoniidae sp NS PC024 G480 LN6258 Tyrannochthonius sp S3 PC025 IS11 PC007 IV203 DD14MRR0004T1 04 PC011 0.72 0.90 0.97 IV279 127105 Pseudoscopiones PC048 1.00 IV278 127104 Pseudoscopiones PC051 0.54 IV210 MEARC3814 20150606 02sc PC012 1.00 1.00 IV269 RC15MEB0171 20160317 T3 01 PC050 1.00 IV268 RC15MEB0171 20160317 T2 02 PC049 JU05 RC15MEH0315.20160627.T2 02 PC014 1.00 JU07 RC15MEH0335.20160627.T1 01 PC015 0.77 IS10 KBRCopp15-20150913-02 PC006 0.78 IV261 GR15MEB0022 20160115 sc 01 PC047 CM18 RC11KOOD0193P2T1 2 Lagynochthonius sp. PC008 AX82 ID31P1T1 5 PC043 0.58 1.00 AX86 ID32P1T1 7 PC044 G324 101086 Tyrannochthonius sp Mariilana Creek PC028 1.00 EA0270R Lag B2 Ophthalmia Range PC033 EXR1148 J20 6 Lagy S4 Hancock Range North PC039 0.76 0.55 GF0173R Lag B2 Jirrpalpur Range PC040 1.00 JIN0078R BHP772 Lagynochthonius Hancock Range South PC041 0.71 G328 LN9199 Tyrannochthonius sp Ophthalmia Range PC029 0.94 PI012 Tyran Wheelarra Hill PC038 AK14 PE110372 Tyrannochthonius DC4 PC013 0.63 0.99 G319 LN8720 Chthoniid sp Munjina PC027 1.00 EJR0202 2 9 9 ob18 Tyrranochthonius B2 PC036 1.00 0.96 LB084 Tyran Wheelarra Hill PC037 G118 Tyrannochthonius Hamersley Range PC021 1.00 1.00 G428 110726 Chthoniidae sp PC031 G430 100975 Tyrannochthonius sp PC030 AX85 ID32P1T1 7 PC045 EU559513 Austrochtonia sp

0.1

21 Molecular systematics of subfauna from the Robe Valley

Figure 4. Bayesian analysis of COI haplotypes of Hyiidae and Olpiidae (Pseudoscorpiones) from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site. AY156582 Pandinus imperator DQ513111 Siro rubens 127103 Pseudoscorpiones PH019 1.00 IV202 DD14MRR0004T1 04 Pseudoscorpiones PH005 IT72 BHRC0512 151013 T3 01 Pseudoscorpiones PH018 0.78 IV213 DD14MRR0009 20151002 T2 02 Pseudoscorpiones PH007 1.00 1.00 0.81 JF56 RC14MEH0252 20160120 T1 04 Pseudoscorpiones PH017 IV211 RC13MEA0279 20150807 T1 02 Pseudoscorpiones PH006 IV216 138532 Pseudoscorpiones PH024 1.00 1.00 IV222 138537 Pseudoscorpiones PH025 1.00 IV219 RC14MEB0101 20151001 T1 02 Pseudoscorpiones PH008 0.97 0.95 IV224 138538 Pseudoscorpiones PH022 0.84 IV259 DD15MEB0018 20160317 T3 01 Pseudoscorpiones PH023 1.00 IV221 139896 Pseudoscorpiones PH020 IV265 RC15MEB0020 20160317 T2 02 Pseudoscorpiones PH021 Hyiidae AX81 ID27P1T4 5 Pseudoscorpiones PH013 0.98 G211 Pseudoscorpiones 109256 PH011 FZ1 Pseudoscorpiones RC13KOOD0302P7T1 1 PH015 1.00 0.98 1.00 1.00 FZ2 Pseudoscorpiones RC13KOOD0305P7T3 3 PH015 JD01 RC13KOOD0184 151011 T1 03 Pseudoscorpiones PH016 GB0018R Jirrpalpur Range Hyiidae B1 PH009 0.99 1.00 1.00 GC0103R Jirrpalpur Range Hyiidae PH009

0.91 PSF0090R LN2124 Packsaddle Indohya PH010 1.00 IV201 DD14MEB0002 20150607 01sc Pseudoscorpiones PH001 1.00 IV217 RC14MEB0029 20151001 T1 01 Pseudoscorpiones PH002 EU559564 Indohya sp. JM 2008 DF18 8668 Indohya sp PH014. 1.00 G107 Indohya PH012 1.00 IV206 DD14MRR0008 20150808 T1 02 Pseudoscorpiones PH003 1.00 IV209 M2ERC027 t2 03 Pseudoscorpiones PH004

0.99 BQ20 PES 1896 Beierolpium won1 PO010 0.97 EU559545 Beierolpium bornemisszai Olpiidae 0.52 IS03 Nolabel-20150709-T1-02 PO011 0.99 IV277 99264 Pseudoscopiones PO012 1.00 0.69 JF59 RC15MEH0302 20151210 T1 03 Pseudoscorpiones PO008 EU559544 Calocheiridius termitophilus 0.79 JD02 RC12KOOD2579 151010 T4 02 Pseudoscorpiones PO009 G320 LN8818 Olpiidae sp Munjina PO003 1.00 0.70 Olpiidae G326 LN7150 Olpiidae sp Marillana Ck PO004 0.60 YYAC26 G89 LN7692 Yeelirrie Station PO001 0.86 0.97 YYHC0048L G90 LN7592 Yeelirrie Station PO002 G322 100795 Olpiidae sp Munjina PO005 0.60 1.00 IV281 132672 Pseudoscopiones PO013 G323 100292 Olpiidae sp MJ PO006 0.72 G325 LN9467 Olpiidae sp MA PO007

0.1

22 Molecular systematics of subfauna from the Robe Valley

Figure 5. Neighbour-joining analysis of specimens of Schizomida from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

IV291 110830 Schizomid IV329 RC14MEB0102-20160317-T3-06 Schizomid IV290 133445 Schizomid IV289 110827 Schizomid IV287 133444 Schizomid IV286 92222 Schizomid 94 1 IV327 DD15MEB0018-20160317-T3-02 Schizomid 63 IV292 140989 Schizomid IV284 65761 Schizomid 100 57 IV328 GR15MEB0004-20160317-T1-01 Schizomid 57 IV330 RC15MEB0020-20160317-T3-02 Schizomid 100 IV288 110826 Schizomid IV295 133446 Schizomid 2 99 100 IV293 140982 Schizomid 3 IV294 140983 Schizomid IV326 DD15MEB0008-20160317-T2-01 Schizomid 4

100 WAMT137066 Bamazomus subsolanus 5 WAMT137067 Bamazomus subsolanus

0.02

23 Molecular systematics of subfauna from the Robe Valley

Figure 6. Bayesian analysis of COI haplotypes of Schizomida from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

EU272675 Brignolozomus woodwardi

IV289 110827 Schizomid SCH001 1.00 0.67 IV61 DD14MEC0002 20150929 T3 03 SCH001 1.00 WAM T63344 Paradraculoides bythius IV07 Budgie20150604 06 SCH002 0.74 IV295 133446 Schizomid SCH036 1.00 1.00 WAM T66236 Paradraculoides anachoretus 1.00 IV293 140982 Schizomid SCH034 IV82 TOBRC0023 20151001 T1 01 SCH003 0.62 1.00 Paradraculoides sp. OFB 2007 WAM T63371 3 Paradraculoides gnophicola 1.00 WAM T65802 Paradraculoides kryptus

1.00 IV76 MELUNK11 20151001 T2 01 SCH004 IV09 DD14MEL0001 t2 01 SCH004a C119 ex T95543 SCH018 0.80 1.00 GH7 Draculoides Dra PES16433 SCH026 1.00 GH8 Draculoides Dra PES16447 SCH027 C37 T92541B SCH016 0.98 HE17 BHRC151 20141216 T3 01c SCH028 0.93 0.90 HE4 BHRC001 20141028 SC01 SCH029 0.57 0.85 IS33 Kbrc116720150809t2 05 Schizomida sp SCH031 IS15 KBRC1389 20150909 04 Schizomida SCH032 0.89 0.99 0.92 IT67 BHRC401 151013 T1 01 Schizomida SCH033 C58 T98320 SCH017 0.92 T93233 C17 CBRC099P5T1 3 Paradraculoides ‘cardo’ IV17 DD14MRR007 20150808 T1 02 SCH006 1.00 IV14 DD14MRR0005 T1 01 SCH007 1.00 IV27 M2ERC0053 T2 02 SCH008 0.92 1.00 1.00 EU272730 Draculoides mesozeirus

1.00 IV55 RC13MRR0077 T1 02 SCH009 U72 Shizomid T95463 SCH019 1.00 U74 Shizomid T95462 SCH020 0.63 1.00 U83 Shizomid T95450 SCH021 EU272684 Draculoides bramstokeri IV81 RC14MRR0017 20151002 T2 01 SCH0010 FT4 Draculoides SCH012 PSB0166R SCH022 0.95 0.72 0.55 1.00 FT6 Draculoides SCH012 GBP553 SCH023 1.00 GG9 Draculoides sp PSA0806R SCH030

1.00 FT5 Draculoides sp PSB0163R SCH024 FZ21 Schizomida RC13KOOD0308P7T1 1 SCH025 IV326 DD15MEB0008 20160317 T2 01 Schizomid SCH005 1.00 0.95 IV79 RC14MEB0060 20151001 T2 01 SCH005 WAMT137066 Bamazomus subsolanus SCH035 0.1

24 Molecular systematics of subfauna from the Robe Valley

Figure 7. Bayesian analysis of COI haplotypes of Thysanura from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site.

JQ029134 Palomena prasina KM013315 Graphosoma lineatum Atel EU084035 Atelura formicaria Atel EA0118R Ophthalmia TA001 Atel BQ13 PES 4627 Atelurinae won1 TA008

1.00 Atel A32 Cape Preston TA007 0.56 1.00 Atel NB1 Cape Preston TA007 Atel IV228 RC14MEB0060 TA009 0.81 1.00 0.98 Atel IV229 RC14MEB0060 TA009 1.000.99 Atel EEX0078 Hancock Range S. TA010 Atel WRKRC148 J7 7 TA011

0.640.68 Atel BQ14 PES 5086 Atelurinae won1 TA002 Atel GA0011R Jirrpalpur Range TA003 1.00 Atel EXR1139 Alligator Range TA004 1.00 Atel EXR1386 J8 14 Mudlark TA004 1.00 Atel G313 101035 Ateluridae MA TA005 Atel BX3 PES 5124 Atelurinae MH1 TA006 Nic JQ282164 Trinemura callawa CA0006 1.00 0.75 Nic JQ282165 Trinemura cundalin CU0060R Nic G318 LN8588 Nicoletiidae sp. MC TN001 1.00 0.77 Nic AY2 BES16311 B10 TN005 Nic PROP003 J16 2 Trimenura TN002 1.00 Nic BX43 PES 5964 Trinemura MH1 TN007 1.00 Nic G126 TN003 0.99 Nic G206 108042 TN004 1.00 0.99 Nic CQ11 Trinemura sp. B15 CC0179 TN009 0.52 0.62 0.98 Nic CQ13 Trinemura sp. B19 CC0748 TN016 Nic EJ0245R J2 36 OB18 TN011 Nic BX42 PES 5266 Trinemura MH1 TN006 Nic CQ10 Trinemura sp. B18 Christmas Ck. TN008 0.99 Nic IV338 GR15MEC0001 20160316 T2 06 TN012 0.97 1.00 Nic IV339 RC15MEC0197 20160316 T2 01 TN012 1.00 0.77 Nic FL3 TOBRC0011PIT2 1 A Barrow Island TN014 1.00 1.00 Nic IV227 MEARC4383 TN010 0.901.00 Nic IV333 MEBRC0020T1 20 TN010

1.00 Nic EXR0983 BHP821 J8 13 Wheelarra Hill TN015 Nic IV334 M2ERC0057T1 21 TN011 Nic CA35 11 1513 Trinemura indet. TN013 Thys IV337 TOBRC0055P1T1 1 T001 1.00 Thys IV344 RC15MEB0171 20150115 SC 01 T002 1.00 Thys IV331 MEGRC130P1T1 1 T003 1.00 Thys IV340 GR15MEB0003 20150115 SC 01 T003 1.00 Thys IV341 GR15MEB0009 20150115 SC 01 T003 Thys IV342 GR15MEB0014 Sc 02 T003 Subnic KCRC0107 TS001

1.00 Subnic FH4 Hemitrinemura sp B10 TM0087 TS002 0.94 Subnic IV230 MEARC3814 TS004 1.00 0.98 Subnic GH1 PES14085 TS004 Subnic FH5 Hemitrinemura sp B10 BG0082 TS003 0.1

25 Table 2. Estimates of Evolutionary Divergence between Sequences

Specimen ID KD01_GR15MEB0014.20160915.T1-01_Aeaneae KD02_RC15MEC0001.20160915.T1-01_Araneae IV346_SSp-2010-75_DCBRC_040_Araneae IV345_SSp-2010-253_DCBRC_017_Araneae IV276_138441_Araneae IV275_127039_Araneae IV272_MEBRC0016P1T1-2_Araneae EF050286_Coleosoma_acutiventer_Theriidae KX537094_Theridion_boesenbergi_Theriidae KR864743_Ischnothyreus_auritus_Oonopidae KJ957966_Gea_theridioides_Aranidae KJ957946_Arachnura_scorpionoides_Aranidae GU684645_Theridion_ohlerti_Theridiidae GH12_Selinopidae_PES16307 G309_LN8606_Prethopalpus_sp_MA G308_LN9827_Oonopidae_sp_MJ G307_LN9925_Gnaphosidae_sp_MJ G227_Araneae_MC8638 G226_Araneae_107786 G210_Araneae_107945 G209_Araneae_109675 G145_Desognanops_sp._n._Y1_LN6635_x2 G142_Desognanops_sp._n._Y1_LN8482 G141_Prethopalpus_sp._S2_LN8445_x2 G140_Oonopidae_sp._indet._LN8185 G139_Oonopidae_sp._indet._LN8432 G137_Oonopidae_sp._indet._100075 G136_Oonopidae_sp._indet._LN7327_x2 G135_Prethopalpus_sp._S2_LN7377_x2 FG1100_BHP877_Jimblebar EO8_Prethopalpus_sp_EMP0049 EO7_Prethopalpus_sp_EMP0061_x2 EO33_Prethopalpus_sp_ECP0057 EJ0003R_OB18 DF9_8999_Araneomorphae_sp. DF8_8662_Anapistula_sp. CA6_11:1510_Araneae_indet. BX1_PES-5149_Anapistula_MH1_Symphytognathidae BP013_Deposit_B_x2 AY231053_Robertus_neglectus_Theriidae KD01_GR15MEB0014.20160915.T1-01_Araneae 0.004 0.022 0.022 0.020 0.015 0.017 0.013 0.013 0.021 0.015 0.018 0.015 0.017 0.020 0.019 0.020 0.020 0.021 0.021 0.019 0.018 0.019 0.019 0.021 0.018 0.018 0.020 0.020 0.017 0.020 0.025 0.024 0.017 0.020 0.020 0.015 0.018 0.015 0.015 KD02_RC15MEC0001.20160915.T1-01_Araneae 0.009 0.021 0.021 0.020 0.015 0.017 0.012 0.013 0.021 0.015 0.018 0.015 0.017 0.019 0.018 0.019 0.020 0.021 0.021 0.019 0.019 0.019 0.018 0.022 0.018 0.018 0.021 0.020 0.017 0.020 0.025 0.024 0.017 0.019 0.020 0.016 0.018 0.014 0.015 IV346_SSp-2010-75_DCBRC_040_Araneae 0.218 0.219 0.005 0.022 0.021 0.017 0.020 0.023 0.019 0.021 0.021 0.022 0.020 0.015 0.018 0.018 0.020 0.019 0.020 0.021 0.018 0.019 0.017 0.018 0.018 0.017 0.017 0.018 0.020 0.018 0.020 0.018 0.020 0.018 0.020 0.017 0.021 0.021 0.022 IV345_SSp-2010-253_DCBRC_017_Araneae 0.216 0.217 0.014 0.022 0.021 0.017 0.019 0.023 0.020 0.021 0.021 0.021 0.020 0.015 0.018 0.018 0.020 0.020 0.020 0.022 0.018 0.020 0.017 0.019 0.018 0.016 0.017 0.018 0.020 0.018 0.020 0.018 0.020 0.018 0.021 0.017 0.022 0.021 0.021 IV276_138441_Araneae 0.225 0.221 0.266 0.269 0.021 0.019 0.020 0.021 0.021 0.021 0.021 0.020 0.019 0.023 0.022 0.022 0.016 0.017 0.017 0.016 0.020 0.021 0.019 0.021 0.021 0.020 0.020 0.020 0.019 0.021 0.023 0.023 0.020 0.019 0.015 0.020 0.014 0.021 0.021 IV275_127039_Araneae 0.174 0.175 0.242 0.242 0.230 0.020 0.012 0.014 0.020 0.015 0.014 0.016 0.015 0.020 0.019 0.020 0.019 0.020 0.019 0.019 0.017 0.017 0.017 0.019 0.018 0.018 0.018 0.019 0.017 0.021 0.026 0.024 0.017 0.019 0.018 0.016 0.018 0.015 0.014 IV272_MEBRC0016P1T1-2_Araneae 0.221 0.218 0.171 0.178 0.250 0.240 0.018 0.020 0.017 0.019 0.019 0.020 0.016 0.018 0.018 0.018 0.019 0.018 0.018 0.020 0.017 0.018 0.017 0.018 0.017 0.017 0.017 0.018 0.019 0.022 0.021 0.020 0.018 0.018 0.017 0.016 0.018 0.019 0.018 EF050286_Coleosoma_acutiventer_Theriidae 0.096 0.093 0.219 0.221 0.210 0.141 0.234 0.012 0.021 0.012 0.014 0.014 0.015 0.018 0.018 0.020 0.018 0.019 0.020 0.019 0.016 0.016 0.017 0.019 0.017 0.017 0.019 0.019 0.016 0.020 0.024 0.022 0.016 0.018 0.018 0.015 0.019 0.015 0.013 KX537094_Theridion_boesenbergi_Theriidae 0.114 0.111 0.239 0.236 0.245 0.136 0.257 0.088 0.020 0.015 0.015 0.015 0.017 0.019 0.021 0.022 0.019 0.021 0.021 0.019 0.018 0.018 0.019 0.021 0.019 0.020 0.021 0.021 0.017 0.022 0.026 0.023 0.017 0.019 0.020 0.017 0.019 0.016 0.013 KR864743_Ischnothyreus_auritus_Oonopidae 0.199 0.204 0.218 0.220 0.279 0.236 0.164 0.245 0.246 0.019 0.019 0.022 0.018 0.019 0.018 0.017 0.020 0.020 0.020 0.020 0.018 0.020 0.017 0.018 0.018 0.018 0.018 0.017 0.020 0.020 0.022 0.022 0.019 0.020 0.020 0.016 0.020 0.019 0.019 KJ957966_Gea_theridioides_Aranidae 0.163 0.164 0.226 0.226 0.235 0.124 0.240 0.137 0.150 0.245 0.014 0.013 0.017 0.019 0.017 0.017 0.018 0.019 0.019 0.018 0.018 0.017 0.018 0.019 0.020 0.019 0.021 0.020 0.016 0.019 0.027 0.025 0.016 0.019 0.016 0.020 0.018 0.017 0.016 KJ957946_Arachnura_scorpionoides_Aranidae 0.174 0.172 0.224 0.228 0.230 0.114 0.220 0.155 0.156 0.241 0.140 0.017 0.017 0.019 0.017 0.018 0.019 0.019 0.019 0.018 0.017 0.017 0.020 0.019 0.020 0.020 0.019 0.020 0.018 0.021 0.026 0.024 0.018 0.019 0.020 0.019 0.020 0.015 0.015 GU684645_Theridion_ohlerti_Theridiidae 0.123 0.123 0.241 0.236 0.186 0.147 0.248 0.101 0.125 0.254 0.136 0.147 0.018 0.018 0.018 0.020 0.019 0.020 0.020 0.020 0.017 0.020 0.018 0.020 0.019 0.019 0.019 0.019 0.016 0.019 0.024 0.023 0.016 0.020 0.019 0.017 0.018 0.015 0.015 GH12_Selinopidae_PES16307 0.178 0.177 0.219 0.221 0.282 0.198 0.242 0.166 0.161 0.223 0.188 0.200 0.192 0.018 0.019 0.018 0.018 0.019 0.019 0.019 0.015 0.017 0.018 0.019 0.019 0.018 0.019 0.020 0.016 0.017 0.025 0.021 0.015 0.017 0.017 0.018 0.019 0.017 0.015 G309_LN8606_Prethopalpus_sp_MA 0.211 0.216 0.169 0.174 0.266 0.246 0.211 0.195 0.225 0.227 0.226 0.213 0.220 0.218 0.015 0.015 0.020 0.021 0.021 0.019 0.020 0.019 0.018 0.018 0.017 0.018 0.017 0.017 0.019 0.017 0.021 0.019 0.020 0.018 0.021 0.017 0.020 0.022 0.019 G308_LN9827_Oonopidae_sp_MJ 0.202 0.203 0.176 0.176 0.261 0.224 0.202 0.206 0.217 0.225 0.232 0.204 0.227 0.220 0.173 0.010 0.019 0.020 0.020 0.020 0.019 0.020 0.019 0.015 0.016 0.016 0.016 0.016 0.016 0.019 0.019 0.018 0.016 0.015 0.021 0.017 0.021 0.020 0.018 G307_LN9925_Gnaphosidae_sp_MJ 0.218 0.212 0.169 0.171 0.269 0.230 0.206 0.228 0.227 0.218 0.224 0.213 0.246 0.218 0.171 0.059 0.019 0.020 0.021 0.020 0.020 0.019 0.019 0.017 0.017 0.018 0.017 0.017 0.017 0.018 0.020 0.019 0.018 0.016 0.021 0.017 0.020 0.020 0.019 G227_Araneae_MC8638 0.212 0.204 0.235 0.235 0.132 0.200 0.234 0.198 0.214 0.255 0.217 0.202 0.209 0.242 0.237 0.226 0.226 0.018 0.019 0.017 0.020 0.019 0.021 0.020 0.020 0.018 0.019 0.019 0.018 0.019 0.022 0.021 0.019 0.018 0.017 0.020 0.015 0.019 0.019 G226_Araneae_107786 0.216 0.212 0.248 0.253 0.124 0.239 0.230 0.219 0.228 0.253 0.241 0.232 0.214 0.244 0.255 0.244 0.248 0.134 0.007 0.018 0.019 0.020 0.021 0.022 0.022 0.022 0.021 0.021 0.020 0.020 0.025 0.023 0.019 0.020 0.014 0.020 0.017 0.021 0.020 G210_Araneae_107945 0.220 0.216 0.241 0.245 0.122 0.239 0.236 0.220 0.232 0.252 0.245 0.232 0.221 0.250 0.264 0.252 0.250 0.135 0.025 0.019 0.018 0.019 0.021 0.022 0.022 0.022 0.022 0.022 0.020 0.020 0.025 0.023 0.020 0.021 0.015 0.021 0.018 0.020 0.019 G209_Araneae_109675 0.210 0.216 0.251 0.249 0.140 0.232 0.245 0.215 0.230 0.253 0.232 0.223 0.210 0.268 0.246 0.218 0.222 0.133 0.144 0.136 0.019 0.019 0.021 0.018 0.021 0.020 0.019 0.019 0.019 0.021 0.025 0.023 0.019 0.019 0.014 0.019 0.017 0.019 0.020 G145_Desognanops_sp._n._Y1_LN6635_x2 0.181 0.180 0.203 0.203 0.241 0.198 0.213 0.172 0.203 0.230 0.221 0.200 0.189 0.182 0.195 0.206 0.213 0.213 0.231 0.230 0.240 0.011 0.017 0.019 0.018 0.017 0.018 0.018 0.016 0.019 0.024 0.022 0.016 0.018 0.020 0.017 0.020 0.018 0.017 G142_Desognanops_sp._n._Y1_LN8482 0.177 0.176 0.192 0.192 0.256 0.183 0.202 0.181 0.189 0.246 0.204 0.191 0.194 0.189 0.215 0.191 0.208 0.203 0.235 0.246 0.240 0.085 0.018 0.018 0.018 0.018 0.017 0.018 0.017 0.019 0.025 0.022 0.017 0.017 0.020 0.019 0.021 0.018 0.016 G141_Prethopalpus_sp._S2_LN8445_x2 0.226 0.227 0.178 0.187 0.269 0.226 0.200 0.213 0.247 0.226 0.240 0.251 0.249 0.232 0.173 0.173 0.182 0.247 0.269 0.278 0.255 0.202 0.202 0.015 0.016 0.015 0.014 0.015 0.020 0.021 0.025 0.022 0.020 0.015 0.022 0.016 0.021 0.019 0.017 G140_Oonopidae_sp._indet._LN8185 0.228 0.224 0.187 0.189 0.274 0.234 0.189 0.215 0.233 0.205 0.253 0.217 0.240 0.221 0.184 0.182 0.182 0.230 0.283 0.278 0.236 0.209 0.204 0.123 0.010 0.011 0.009 0.009 0.019 0.020 0.023 0.021 0.019 0.015 0.020 0.015 0.021 0.020 0.018 G139_Oonopidae_sp._indet._LN8432 0.226 0.222 0.212 0.217 0.289 0.238 0.202 0.213 0.230 0.221 0.251 0.230 0.240 0.223 0.169 0.178 0.182 0.254 0.295 0.283 0.249 0.204 0.211 0.130 0.049 0.010 0.009 0.009 0.020 0.019 0.025 0.021 0.019 0.015 0.019 0.015 0.020 0.021 0.018 G137_Oonopidae_sp._indet._100075 0.215 0.211 0.192 0.192 0.264 0.232 0.191 0.206 0.221 0.210 0.247 0.226 0.233 0.223 0.184 0.169 0.178 0.235 0.281 0.278 0.249 0.191 0.209 0.130 0.060 0.053 0.011 0.010 0.019 0.021 0.024 0.021 0.019 0.016 0.020 0.015 0.021 0.021 0.018 G136_Oonopidae_sp._indet._LN7327_x2 0.230 0.227 0.194 0.199 0.274 0.230 0.191 0.215 0.233 0.199 0.257 0.223 0.242 0.212 0.182 0.178 0.180 0.232 0.288 0.276 0.253 0.191 0.209 0.130 0.036 0.043 0.057 0.006 0.019 0.019 0.024 0.020 0.019 0.015 0.020 0.015 0.021 0.021 0.018 G135_Prethopalpus_sp._S2_LN7377_x2 0.226 0.222 0.194 0.199 0.272 0.232 0.194 0.213 0.233 0.199 0.251 0.223 0.235 0.210 0.173 0.184 0.182 0.232 0.285 0.276 0.245 0.187 0.215 0.132 0.030 0.040 0.055 0.019 0.020 0.020 0.023 0.020 0.020 0.016 0.020 0.015 0.021 0.021 0.018 FG1100_BHP877_Jimblebar 0.172 0.180 0.221 0.219 0.237 0.175 0.224 0.169 0.193 0.231 0.167 0.171 0.173 0.180 0.206 0.224 0.246 0.212 0.228 0.225 0.214 0.180 0.199 0.227 0.208 0.223 0.212 0.219 0.219 0.020 0.025 0.023 0.004 0.019 0.018 0.017 0.019 0.015 0.017 EO8_Prethopalpus_sp_EMP0049 0.230 0.232 0.144 0.154 0.265 0.260 0.210 0.263 0.258 0.230 0.245 0.240 0.253 0.242 0.189 0.187 0.192 0.240 0.240 0.235 0.260 0.237 0.215 0.192 0.189 0.199 0.207 0.194 0.189 0.245 0.021 0.019 0.020 0.018 0.020 0.020 0.020 0.021 0.020 EO7_Prethopalpus_sp_EMP0061_x2 0.285 0.282 0.176 0.189 0.279 0.303 0.211 0.279 0.294 0.257 0.297 0.294 0.297 0.276 0.220 0.167 0.211 0.260 0.272 0.260 0.285 0.282 0.282 0.189 0.192 0.211 0.201 0.201 0.201 0.257 0.149 0.011 0.025 0.022 0.023 0.022 0.025 0.026 0.025 EO33_Prethopalpus_sp_ECP0057 0.273 0.264 0.179 0.190 0.261 0.293 0.207 0.276 0.281 0.256 0.287 0.287 0.290 0.278 0.210 0.170 0.199 0.253 0.264 0.247 0.267 0.267 0.276 0.188 0.188 0.190 0.182 0.179 0.185 0.259 0.145 0.038 0.023 0.019 0.021 0.022 0.021 0.025 0.023 EJ0003R_OB18 0.168 0.174 0.219 0.217 0.238 0.174 0.221 0.166 0.189 0.230 0.166 0.170 0.169 0.172 0.209 0.231 0.249 0.216 0.226 0.227 0.219 0.182 0.202 0.230 0.210 0.226 0.215 0.221 0.221 0.008 0.245 0.257 0.259 0.019 0.017 0.017 0.019 0.014 0.017 DF9_8999_Araneomorphae_sp. 0.228 0.231 0.169 0.169 0.314 0.252 0.211 0.241 0.239 0.217 0.245 0.234 0.259 0.194 0.198 0.186 0.200 0.261 0.292 0.287 0.266 0.246 0.231 0.184 0.175 0.195 0.191 0.186 0.186 0.227 0.187 0.198 0.207 0.228 0.018 0.016 0.019 0.020 0.017 DF8_8662_Anapistula_sp. 0.210 0.205 0.237 0.237 0.108 0.228 0.232 0.198 0.225 0.254 0.226 0.230 0.196 0.241 0.229 0.214 0.223 0.130 0.105 0.117 0.130 0.235 0.235 0.252 0.252 0.254 0.257 0.254 0.252 0.223 0.245 0.263 0.250 0.215 0.273 0.018 0.015 0.020 0.020 CA6_11:1510_Araneae_indet. 0.215 0.207 0.189 0.194 0.250 0.227 0.222 0.179 0.234 0.208 0.231 0.235 0.223 0.222 0.176 0.189 0.194 0.247 0.253 0.264 0.254 0.214 0.216 0.165 0.169 0.178 0.180 0.185 0.183 0.230 0.217 0.214 0.222 0.224 0.220 0.235 0.019 0.016 0.018 BX1_PES-5149_Anapistula_MH1_Symphytognathidae 0.206 0.205 0.240 0.244 0.115 0.235 0.255 0.203 0.221 0.269 0.235 0.238 0.212 0.233 0.228 0.240 0.240 0.124 0.121 0.135 0.134 0.243 0.243 0.263 0.249 0.263 0.254 0.252 0.245 0.233 0.247 0.272 0.270 0.227 0.279 0.112 0.246 0.020 0.018 BP013_Deposit_B_x2 0.166 0.165 0.231 0.229 0.243 0.163 0.251 0.160 0.169 0.247 0.172 0.168 0.158 0.170 0.205 0.238 0.249 0.223 0.233 0.241 0.233 0.183 0.198 0.224 0.230 0.241 0.235 0.235 0.226 0.130 0.260 0.300 0.284 0.119 0.236 0.230 0.207 0.224 0.015 AY231053_Robertus_neglectus_Theriidae 0.139 0.136 0.235 0.237 0.201 0.133 0.223 0.104 0.118 0.234 0.129 0.127 0.116 0.155 0.215 0.229 0.240 0.196 0.210 0.220 0.228 0.190 0.181 0.226 0.223 0.223 0.226 0.226 0.221 0.177 0.258 0.285 0.284 0.173 0.213 0.210 0.199 0.190 0.152 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 6. Estimates of Evolutionary Divergence between Sequences

Lineage IV261 IV279 IV268 IV269 IV278 An-sp. Ljoh Pa-sp. Tsp Ps-sp2 Au-sp Ps-sp Sins Tari IS01 IS12 IS08 IS06 IS05 IS10 IS11 CM18 IV212 IV226 IV203 IV210 AK14 JU05 JU07 G146 G149 G154 G151 G152 G118 G219 G220 G479 G480 G85 G319 G324 G328 G430 G428 EAP0178 EA0270R G87 J8_31 EJR0202 LB084 PI012 J20-6 GF0173R JIN0078R AX83 AX82 AX86 AX85 JV08 IV261 GR15MEB0022-20160115-sc-01 Pseudoscorpiones Gp 5 PC047 0.015 0.016 0.014 0.015 0.021 0.021 0.018 0.020 0.021 0.019 0.016 0.017 0.020 0.017 0.017 0.018 0.020 0.019 0.018 0.019 0.018 0.017 0.019 0.018 0.019 0.017 0.014 0.015 0.018 0.019 0.018 0.020 0.019 0.024 0.021 0.021 0.018 0.016 0.017 0.021 0.019 0.018 0.016 0.015 0.026 0.025 0.019 0.026 0.024 0.022 0.022 0.025 0.023 0.024 0.015 0.015 0.018 0.014 0.017 IV279 127105 Pseudoscopiones Gp 4 PC048 0.178 0.015 0.015 0.011 0.021 0.020 0.020 0.020 0.023 0.020 0.017 0.016 0.020 0.015 0.015 0.014 0.019 0.020 0.017 0.018 0.021 0.018 0.018 0.016 0.017 0.021 0.014 0.015 0.020 0.020 0.020 0.019 0.018 0.024 0.022 0.020 0.016 0.015 0.019 0.020 0.017 0.020 0.017 0.015 0.023 0.023 0.018 0.021 0.021 0.021 0.024 0.024 0.020 0.024 0.015 0.017 0.018 0.016 0.018 IV268 RC15MEB0171-20160317-T2-02 Pseudoscorpiones Gp 1 PC049 0.212 0.175 0.011 0.014 0.020 0.020 0.019 0.021 0.021 0.020 0.018 0.016 0.018 0.017 0.017 0.016 0.017 0.016 0.017 0.018 0.020 0.018 0.019 0.021 0.014 0.022 0.014 0.014 0.020 0.020 0.019 0.018 0.019 0.023 0.018 0.018 0.015 0.014 0.019 0.022 0.019 0.019 0.014 0.015 0.022 0.023 0.021 0.021 0.021 0.020 0.020 0.024 0.024 0.023 0.014 0.014 0.016 0.013 0.017 IV269 RC15MEB0171-20160317-T3-01 Pseudoscorpiones Gp 2 PC050 0.186 0.175 0.117 0.014 0.021 0.020 0.019 0.020 0.021 0.019 0.015 0.016 0.019 0.017 0.018 0.018 0.017 0.017 0.019 0.017 0.018 0.018 0.019 0.019 0.013 0.021 0.013 0.014 0.018 0.019 0.018 0.019 0.018 0.023 0.019 0.018 0.015 0.014 0.019 0.023 0.017 0.017 0.012 0.014 0.023 0.022 0.019 0.022 0.021 0.022 0.022 0.024 0.022 0.022 0.014 0.014 0.015 0.014 0.016 IV278 127104 Pseudoscopiones Gp 3 PC051 0.193 0.104 0.165 0.154 0.019 0.021 0.018 0.019 0.022 0.019 0.015 0.018 0.021 0.017 0.017 0.015 0.016 0.017 0.018 0.020 0.022 0.019 0.019 0.015 0.015 0.019 0.014 0.014 0.019 0.019 0.017 0.019 0.017 0.024 0.020 0.020 0.016 0.015 0.016 0.019 0.017 0.018 0.016 0.016 0.022 0.023 0.019 0.021 0.023 0.020 0.023 0.021 0.022 0.022 0.015 0.015 0.017 0.015 0.017 Anaulacodithella sp. EU559520 0.234 0.216 0.225 0.241 0.216 0.023 0.020 0.019 0.021 0.020 0.023 0.019 0.019 0.022 0.021 0.020 0.023 0.020 0.021 0.020 0.020 0.019 0.022 0.022 0.020 0.019 0.019 0.019 0.019 0.018 0.020 0.018 0.020 0.025 0.021 0.021 0.023 0.020 0.020 0.022 0.021 0.021 0.019 0.020 0.026 0.024 0.021 0.022 0.024 0.020 0.022 0.021 0.023 0.023 0.019 0.020 0.022 0.020 0.020 EU559503 Lagynochthonius johni 0.206 0.212 0.236 0.236 0.238 0.236 0.021 0.022 0.020 0.021 0.023 0.021 0.020 0.021 0.021 0.021 0.022 0.021 0.021 0.019 0.020 0.018 0.020 0.020 0.021 0.021 0.020 0.019 0.020 0.019 0.022 0.020 0.022 0.023 0.021 0.020 0.021 0.020 0.021 0.023 0.022 0.022 0.020 0.020 0.025 0.025 0.021 0.025 0.023 0.025 0.025 0.022 0.024 0.025 0.019 0.024 0.021 0.022 0.020 EU559505 Paraliochthonius sp. JM-2008 0.192 0.172 0.194 0.203 0.201 0.191 0.198 0.017 0.021 0.017 0.023 0.019 0.019 0.018 0.018 0.018 0.020 0.019 0.019 0.016 0.018 0.017 0.018 0.018 0.019 0.019 0.017 0.017 0.019 0.019 0.019 0.018 0.019 0.025 0.017 0.019 0.019 0.017 0.019 0.023 0.018 0.017 0.020 0.019 0.024 0.022 0.019 0.023 0.021 0.019 0.019 0.021 0.021 0.022 0.018 0.017 0.021 0.016 0.019 EU559506 Tyrannochthonius sp. JM-2008 0.238 0.226 0.222 0.226 0.224 0.199 0.229 0.189 0.020 0.017 0.021 0.019 0.019 0.020 0.020 0.019 0.020 0.019 0.017 0.018 0.020 0.019 0.020 0.018 0.018 0.019 0.019 0.018 0.021 0.019 0.019 0.020 0.019 0.024 0.018 0.018 0.020 0.018 0.021 0.025 0.020 0.019 0.019 0.020 0.026 0.022 0.020 0.026 0.023 0.022 0.022 0.023 0.024 0.023 0.019 0.019 0.021 0.020 0.020 EU559509 Pseudotyrannochthonius sp. JM-2 0.238 0.233 0.263 0.254 0.245 0.215 0.229 0.219 0.238 0.020 0.023 0.020 0.023 0.020 0.022 0.021 0.024 0.021 0.023 0.022 0.022 0.021 0.024 0.020 0.019 0.018 0.021 0.022 0.021 0.021 0.022 0.021 0.019 0.023 0.022 0.022 0.021 0.020 0.022 0.022 0.022 0.024 0.021 0.019 0.024 0.025 0.021 0.025 0.022 0.024 0.026 0.024 0.024 0.025 0.021 0.023 0.024 0.021 0.020 EU559513 Austrochtonia sp 0.199 0.189 0.219 0.194 0.192 0.182 0.203 0.161 0.171 0.212 0.018 0.019 0.021 0.017 0.016 0.018 0.021 0.020 0.020 0.019 0.020 0.017 0.020 0.017 0.017 0.017 0.018 0.018 0.017 0.016 0.019 0.021 0.019 0.025 0.019 0.018 0.020 0.017 0.018 0.021 0.017 0.017 0.018 0.021 0.024 0.021 0.018 0.025 0.023 0.023 0.020 0.023 0.021 0.020 0.018 0.018 0.020 0.019 0.021 EU559519 Pseudochthonius sp. 0.216 0.240 0.234 0.219 0.208 0.228 0.258 0.243 0.243 0.275 0.207 0.018 0.021 0.020 0.019 0.020 0.022 0.021 0.021 0.019 0.023 0.021 0.021 0.021 0.019 0.019 0.019 0.019 0.021 0.023 0.020 0.022 0.020 0.025 0.023 0.023 0.019 0.017 0.022 0.021 0.021 0.021 0.017 0.016 0.024 0.025 0.024 0.022 0.022 0.024 0.025 0.022 0.022 0.022 0.017 0.017 0.017 0.017 0.020 EU559521 Sathrochthonius insulanus 0.249 0.240 0.257 0.239 0.233 0.193 0.233 0.231 0.226 0.245 0.182 0.230 0.023 0.020 0.020 0.018 0.023 0.019 0.019 0.020 0.021 0.022 0.020 0.019 0.021 0.018 0.018 0.017 0.020 0.019 0.019 0.020 0.019 0.025 0.021 0.021 0.018 0.018 0.020 0.020 0.021 0.018 0.018 0.019 0.023 0.023 0.018 0.024 0.024 0.022 0.023 0.021 0.022 0.022 0.017 0.018 0.019 0.016 0.019 KJ659959 Tyrannochthonius aridus 0.213 0.191 0.203 0.196 0.191 0.223 0.227 0.191 0.266 0.241 0.202 0.216 0.252 0.020 0.021 0.020 0.023 0.020 0.017 0.018 0.021 0.019 0.020 0.021 0.019 0.021 0.020 0.019 0.018 0.020 0.020 0.018 0.019 0.023 0.021 0.022 0.022 0.020 0.021 0.019 0.018 0.022 0.019 0.020 0.024 0.023 0.021 0.024 0.022 0.021 0.021 0.023 0.022 0.022 0.021 0.019 0.021 0.017 0.019 IS01 PC001 0.225 0.214 0.220 0.209 0.218 0.269 0.249 0.227 0.273 0.247 0.212 0.252 0.253 0.201 0.010 0.014 0.016 0.017 0.017 0.017 0.018 0.019 0.020 0.015 0.016 0.019 0.017 0.017 0.019 0.018 0.018 0.018 0.020 0.023 0.017 0.018 0.018 0.018 0.020 0.019 0.016 0.017 0.017 0.019 0.021 0.020 0.019 0.023 0.024 0.022 0.023 0.023 0.021 0.024 0.020 0.016 0.019 0.017 0.018 IS12 PC002 0.218 0.212 0.232 0.214 0.223 0.264 0.249 0.218 0.270 0.247 0.215 0.245 0.262 0.199 0.053 0.014 0.017 0.016 0.018 0.017 0.017 0.019 0.019 0.016 0.017 0.021 0.017 0.017 0.019 0.020 0.019 0.019 0.021 0.024 0.017 0.019 0.018 0.018 0.020 0.020 0.016 0.017 0.016 0.018 0.020 0.020 0.020 0.023 0.024 0.021 0.023 0.023 0.022 0.024 0.020 0.017 0.019 0.018 0.017 IS08 PC003 0.218 0.216 0.225 0.236 0.216 0.269 0.256 0.232 0.273 0.263 0.229 0.254 0.269 0.211 0.093 0.091 0.015 0.016 0.017 0.015 0.019 0.019 0.019 0.016 0.015 0.020 0.017 0.017 0.018 0.018 0.018 0.018 0.018 0.026 0.017 0.019 0.018 0.017 0.017 0.019 0.017 0.020 0.017 0.018 0.022 0.022 0.019 0.022 0.021 0.021 0.024 0.025 0.022 0.024 0.020 0.016 0.017 0.016 0.017 IS06 PC004 0.238 0.218 0.212 0.220 0.227 0.255 0.229 0.203 0.263 0.254 0.226 0.261 0.257 0.216 0.149 0.147 0.136 0.014 0.019 0.017 0.019 0.018 0.018 0.016 0.017 0.022 0.017 0.016 0.019 0.019 0.018 0.020 0.020 0.024 0.018 0.021 0.020 0.017 0.021 0.021 0.019 0.020 0.019 0.018 0.026 0.022 0.018 0.024 0.024 0.021 0.025 0.023 0.024 0.023 0.017 0.019 0.021 0.018 0.016 IS05 PC005 0.245 0.225 0.218 0.212 0.227 0.260 0.242 0.221 0.268 0.252 0.242 0.261 0.269 0.218 0.167 0.147 0.149 0.116 0.017 0.016 0.018 0.019 0.019 0.018 0.018 0.021 0.019 0.018 0.018 0.018 0.017 0.019 0.019 0.024 0.015 0.020 0.018 0.017 0.020 0.021 0.017 0.018 0.017 0.016 0.022 0.022 0.019 0.022 0.023 0.021 0.025 0.021 0.021 0.024 0.019 0.019 0.018 0.017 0.018 IS10 PC006 0.194 0.192 0.203 0.205 0.187 0.225 0.229 0.191 0.201 0.261 0.208 0.259 0.244 0.235 0.227 0.236 0.222 0.258 0.231 0.017 0.019 0.018 0.020 0.017 0.018 0.020 0.019 0.018 0.021 0.021 0.019 0.019 0.019 0.024 0.019 0.021 0.021 0.018 0.019 0.022 0.020 0.020 0.019 0.017 0.023 0.022 0.019 0.022 0.024 0.022 0.023 0.024 0.025 0.023 0.020 0.020 0.020 0.018 0.018 IS11 PC007 0.185 0.151 0.180 0.194 0.156 0.225 0.210 0.178 0.196 0.233 0.182 0.213 0.228 0.181 0.229 0.233 0.222 0.227 0.209 0.176 0.018 0.019 0.017 0.016 0.017 0.019 0.018 0.018 0.018 0.017 0.018 0.017 0.017 0.027 0.016 0.018 0.017 0.015 0.020 0.019 0.017 0.021 0.018 0.018 0.025 0.024 0.017 0.023 0.024 0.023 0.021 0.024 0.018 0.021 0.018 0.017 0.016 0.016 0.016 CM18 RC11KOOD0193P2T1-2 Lagynochthonius sp. CP008 0.189 0.192 0.236 0.203 0.212 0.241 0.226 0.186 0.233 0.256 0.201 0.245 0.230 0.191 0.238 0.220 0.236 0.222 0.220 0.224 0.189 0.019 0.018 0.019 0.020 0.022 0.020 0.020 0.019 0.019 0.019 0.019 0.020 0.025 0.019 0.019 0.018 0.017 0.019 0.020 0.017 0.018 0.017 0.019 0.024 0.023 0.019 0.027 0.025 0.023 0.023 0.021 0.021 0.021 0.019 0.018 0.018 0.017 0.020 IV212 DD14MEL0001-20151001-T2-02 PC009 0.229 0.209 0.216 0.216 0.212 0.244 0.231 0.221 0.252 0.259 0.194 0.245 0.253 0.169 0.187 0.196 0.189 0.187 0.204 0.240 0.193 0.216 0.015 0.017 0.017 0.020 0.018 0.019 0.018 0.017 0.018 0.017 0.018 0.023 0.019 0.021 0.021 0.020 0.019 0.020 0.017 0.020 0.017 0.017 0.025 0.023 0.020 0.024 0.024 0.021 0.021 0.023 0.021 0.023 0.021 0.017 0.020 0.016 0.019 IV226 RC14MRR0020-20151002-T2-01 PC010 0.227 0.212 0.234 0.227 0.209 0.244 0.226 0.223 0.259 0.254 0.206 0.247 0.230 0.184 0.207 0.202 0.216 0.189 0.202 0.238 0.193 0.216 0.111 0.017 0.020 0.019 0.018 0.020 0.020 0.018 0.019 0.020 0.022 0.025 0.021 0.021 0.020 0.020 0.021 0.022 0.018 0.020 0.019 0.020 0.025 0.023 0.020 0.024 0.025 0.022 0.022 0.023 0.020 0.020 0.019 0.018 0.023 0.018 0.018 IV203 DD14MRR0004T1-04 PC011 0.185 0.122 0.180 0.158 0.125 0.207 0.215 0.167 0.210 0.224 0.164 0.204 0.223 0.184 0.204 0.204 0.216 0.193 0.236 0.200 0.144 0.182 0.204 0.207 0.018 0.018 0.018 0.018 0.017 0.017 0.017 0.020 0.018 0.023 0.020 0.017 0.017 0.017 0.020 0.019 0.016 0.017 0.019 0.018 0.022 0.022 0.017 0.025 0.020 0.022 0.024 0.022 0.021 0.022 0.019 0.020 0.019 0.018 0.017 IV210 MEARC3814-20150606-02sc PC012 0.205 0.163 0.111 0.085 0.143 0.223 0.233 0.205 0.219 0.247 0.187 0.225 0.248 0.208 0.198 0.204 0.207 0.216 0.222 0.193 0.187 0.216 0.218 0.238 0.156 0.021 0.018 0.017 0.018 0.019 0.018 0.017 0.017 0.023 0.020 0.020 0.020 0.018 0.018 0.020 0.018 0.018 0.018 0.017 0.023 0.022 0.018 0.020 0.020 0.018 0.019 0.022 0.023 0.021 0.018 0.018 0.020 0.017 0.017 AK14 PE110372 Tyrannochthonius DC4 PC013 0.193 0.195 0.209 0.205 0.186 0.207 0.233 0.196 0.222 0.229 0.180 0.245 0.214 0.233 0.237 0.246 0.241 0.251 0.248 0.221 0.198 0.211 0.237 0.246 0.172 0.198 0.018 0.018 0.018 0.020 0.019 0.019 0.018 0.025 0.020 0.022 0.022 0.020 0.021 0.018 0.020 0.020 0.021 0.021 0.024 0.021 0.018 0.024 0.024 0.022 0.023 0.023 0.024 0.021 0.019 0.020 0.020 0.018 0.019 JU05 RC15MEH0315.20160627.T2-02 PC014 0.175 0.154 0.161 0.148 0.148 0.221 0.229 0.178 0.219 0.245 0.194 0.236 0.239 0.196 0.200 0.196 0.207 0.211 0.222 0.189 0.198 0.216 0.211 0.224 0.164 0.133 0.191 0.006 0.019 0.018 0.018 0.018 0.017 0.023 0.019 0.020 0.016 0.015 0.017 0.022 0.018 0.018 0.014 0.016 0.022 0.021 0.018 0.023 0.020 0.019 0.022 0.022 0.025 0.025 0.014 0.015 0.018 0.014 0.015 JU07 RC15MEH0335.20160627.T1-01 PC015 0.181 0.160 0.162 0.157 0.157 0.214 0.219 0.173 0.208 0.242 0.192 0.237 0.241 0.186 0.198 0.189 0.204 0.202 0.218 0.187 0.191 0.204 0.204 0.229 0.158 0.133 0.186 0.030 0.019 0.019 0.018 0.018 0.017 0.022 0.019 0.019 0.017 0.015 0.017 0.020 0.018 0.018 0.013 0.017 0.023 0.021 0.018 0.023 0.022 0.019 0.020 0.022 0.025 0.025 0.015 0.017 0.018 0.014 0.015 G146 Pseudoscorpians LN6643 PC016 0.217 0.221 0.217 0.194 0.219 0.212 0.220 0.184 0.208 0.218 0.188 0.204 0.233 0.189 0.216 0.209 0.218 0.214 0.209 0.214 0.203 0.196 0.178 0.196 0.183 0.205 0.207 0.209 0.196 0.009 0.011 0.016 0.017 0.024 0.018 0.020 0.018 0.017 0.018 0.020 0.015 0.018 0.018 0.019 0.025 0.021 0.017 0.024 0.024 0.021 0.021 0.019 0.023 0.022 0.018 0.017 0.019 0.017 0.019 G149 Pseudoscorpians LN8500 PC017 0.230 0.219 0.223 0.199 0.219 0.214 0.211 0.184 0.215 0.229 0.185 0.224 0.233 0.209 0.220 0.218 0.223 0.216 0.209 0.218 0.207 0.207 0.167 0.183 0.187 0.203 0.226 0.209 0.196 0.043 0.012 0.015 0.016 0.024 0.018 0.020 0.018 0.017 0.018 0.020 0.015 0.017 0.017 0.018 0.024 0.022 0.016 0.023 0.022 0.021 0.020 0.019 0.023 0.021 0.018 0.017 0.019 0.017 0.020 G154 Pseduoscorpians LN8374 PC018 0.205 0.207 0.207 0.194 0.220 0.224 0.227 0.187 0.213 0.231 0.190 0.221 0.223 0.209 0.225 0.214 0.209 0.196 0.187 0.198 0.207 0.204 0.187 0.198 0.192 0.192 0.207 0.218 0.204 0.064 0.073 0.016 0.015 0.025 0.018 0.020 0.018 0.016 0.018 0.022 0.017 0.019 0.019 0.018 0.023 0.022 0.016 0.024 0.026 0.022 0.024 0.021 0.024 0.022 0.018 0.020 0.019 0.017 0.019 G151 Pseudoscorpians LN8451 PC019 0.208 0.199 0.214 0.203 0.217 0.210 0.211 0.189 0.204 0.220 0.199 0.233 0.249 0.194 0.214 0.214 0.218 0.209 0.214 0.207 0.194 0.185 0.189 0.198 0.203 0.178 0.233 0.209 0.196 0.119 0.119 0.130 0.015 0.024 0.020 0.021 0.021 0.020 0.018 0.021 0.018 0.020 0.018 0.019 0.024 0.022 0.014 0.024 0.024 0.021 0.022 0.020 0.023 0.023 0.016 0.018 0.020 0.015 0.018 G152 Pseudoscorpians LN7608 PC020 0.212 0.214 0.205 0.189 0.200 0.205 0.206 0.189 0.220 0.208 0.183 0.218 0.232 0.209 0.227 0.227 0.216 0.227 0.238 0.209 0.223 0.228 0.205 0.220 0.178 0.180 0.182 0.191 0.189 0.135 0.126 0.128 0.145 0.027 0.019 0.022 0.019 0.018 0.013 0.022 0.016 0.018 0.018 0.019 0.022 0.022 0.015 0.023 0.025 0.019 0.022 0.020 0.023 0.021 0.017 0.019 0.019 0.018 0.018 G118 Tyrannochthonius Hamersley Range PC021 0.208 0.235 0.243 0.246 0.235 0.273 0.261 0.234 0.255 0.249 0.235 0.270 0.261 0.258 0.258 0.261 0.276 0.282 0.296 0.252 0.270 0.255 0.279 0.287 0.220 0.246 0.249 0.264 0.261 0.246 0.240 0.261 0.270 0.270 0.027 0.024 0.024 0.027 0.024 0.024 0.026 0.025 0.022 0.019 0.025 0.025 0.023 0.025 0.022 0.023 0.023 0.022 0.024 0.028 0.022 0.025 0.024 0.024 0.022 G219 Pseudoscorpian 108935 Turner River PC022 0.191 0.193 0.175 0.166 0.191 0.198 0.226 0.156 0.212 0.224 0.176 0.228 0.232 0.181 0.198 0.186 0.205 0.177 0.168 0.209 0.166 0.198 0.216 0.221 0.184 0.179 0.211 0.161 0.154 0.175 0.184 0.180 0.184 0.191 0.249 0.016 0.017 0.016 0.020 0.022 0.019 0.019 0.017 0.021 0.024 0.019 0.019 0.025 0.021 0.021 0.021 0.021 0.022 0.021 0.018 0.020 0.022 0.018 0.018 G220 Pseudoscorpian 109177 Turner River PC023 0.182 0.184 0.211 0.179 0.186 0.221 0.206 0.189 0.215 0.238 0.185 0.233 0.225 0.203 0.211 0.195 0.214 0.200 0.200 0.230 0.189 0.202 0.211 0.221 0.177 0.195 0.214 0.191 0.179 0.212 0.210 0.200 0.212 0.200 0.243 0.117 0.013 0.012 0.021 0.023 0.020 0.019 0.018 0.019 0.024 0.022 0.020 0.024 0.020 0.023 0.023 0.021 0.021 0.022 0.021 0.020 0.020 0.020 0.018 G479 111446 Chthoniidae sp NS PC024 0.193 0.191 0.200 0.178 0.190 0.225 0.208 0.187 0.217 0.242 0.185 0.242 0.236 0.208 0.203 0.198 0.207 0.192 0.180 0.234 0.174 0.187 0.212 0.220 0.171 0.207 0.223 0.190 0.193 0.208 0.210 0.200 0.205 0.209 0.240 0.124 0.071 0.011 0.020 0.023 0.018 0.019 0.014 0.016 0.022 0.022 0.019 0.026 0.021 0.023 0.022 0.021 0.021 0.022 0.016 0.014 0.015 0.015 0.016 G480 LN6258 Tyrannochthonius sp S3 PC025 0.178 0.182 0.202 0.168 0.180 0.214 0.192 0.174 0.192 0.233 0.164 0.225 0.239 0.199 0.192 0.185 0.198 0.180 0.185 0.207 0.171 0.189 0.212 0.214 0.163 0.187 0.209 0.175 0.173 0.194 0.199 0.187 0.199 0.192 0.238 0.122 0.064 0.063 0.019 0.021 0.018 0.019 0.013 0.016 0.025 0.021 0.017 0.025 0.020 0.021 0.023 0.020 0.021 0.021 0.014 0.014 0.015 0.014 0.016 SB14-MT G85 LN8520 PC026 0.186 0.202 0.215 0.204 0.200 0.218 0.208 0.187 0.229 0.222 0.188 0.232 0.234 0.210 0.242 0.226 0.221 0.239 0.235 0.201 0.208 0.203 0.204 0.210 0.190 0.199 0.211 0.195 0.195 0.136 0.140 0.138 0.147 0.094 0.238 0.183 0.197 0.204 0.188 0.021 0.017 0.018 0.018 0.018 0.023 0.023 0.017 0.025 0.023 0.021 0.023 0.020 0.024 0.022 0.019 0.020 0.021 0.018 0.019 G319 LN8720 Chthoniid sp Munjina PC027 0.232 0.227 0.232 0.235 0.240 0.220 0.235 0.227 0.249 0.264 0.191 0.247 0.252 0.230 0.230 0.232 0.237 0.249 0.267 0.235 0.237 0.242 0.247 0.249 0.210 0.220 0.205 0.237 0.227 0.215 0.218 0.232 0.215 0.225 0.261 0.225 0.237 0.235 0.225 0.227 0.019 0.022 0.023 0.020 0.024 0.022 0.019 0.026 0.024 0.022 0.021 0.023 0.023 0.022 0.017 0.020 0.023 0.019 0.020 G324 101086 Tyrannochthonius sp Mariilana Creek PC028 0.208 0.185 0.220 0.199 0.185 0.222 0.220 0.220 0.232 0.243 0.177 0.222 0.251 0.212 0.210 0.203 0.208 0.217 0.215 0.215 0.187 0.178 0.212 0.212 0.176 0.203 0.206 0.203 0.196 0.178 0.174 0.192 0.187 0.183 0.249 0.187 0.191 0.181 0.178 0.175 0.191 0.017 0.017 0.018 0.022 0.021 0.018 0.023 0.025 0.021 0.022 0.021 0.022 0.022 0.017 0.017 0.020 0.020 0.017 G328 LN9199 Tyrannochthonius sp Ophthalmia Range PC029 0.181 0.183 0.215 0.199 0.190 0.229 0.232 0.188 0.222 0.243 0.175 0.237 0.215 0.194 0.199 0.192 0.224 0.199 0.215 0.217 0.192 0.174 0.203 0.208 0.180 0.194 0.182 0.178 0.176 0.181 0.178 0.192 0.196 0.196 0.217 0.163 0.177 0.183 0.192 0.198 0.205 0.165 0.019 0.020 0.021 0.019 0.019 0.025 0.024 0.020 0.018 0.021 0.020 0.021 0.019 0.018 0.019 0.017 0.019 G430 100975 Tyrannochthonius sp PC030 0.186 0.203 0.198 0.172 0.202 0.207 0.199 0.199 0.222 0.245 0.187 0.237 0.219 0.199 0.256 0.241 0.252 0.236 0.238 0.207 0.178 0.174 0.238 0.234 0.194 0.189 0.214 0.177 0.172 0.232 0.232 0.232 0.208 0.229 0.199 0.186 0.193 0.180 0.178 0.200 0.235 0.172 0.199 0.015 0.025 0.021 0.019 0.024 0.023 0.021 0.021 0.022 0.022 0.023 0.014 0.017 0.018 0.014 0.016 G428 110726 Chthoniidae sp PC031 0.202 0.210 0.218 0.211 0.212 0.218 0.236 0.201 0.247 0.236 0.226 0.263 0.257 0.238 0.234 0.243 0.232 0.216 0.232 0.223 0.234 0.229 0.247 0.238 0.194 0.198 0.241 0.206 0.209 0.221 0.225 0.234 0.230 0.234 0.150 0.216 0.214 0.190 0.191 0.209 0.230 0.217 0.227 0.169 0.025 0.023 0.020 0.024 0.020 0.020 0.022 0.022 0.024 0.026 0.017 0.015 0.016 0.014 0.016 EAP0178 Tyran B1 Ophthalmia Range PC032 0.262 0.259 0.256 0.250 0.256 0.303 0.282 0.264 0.300 0.262 0.241 0.288 0.285 0.235 0.206 0.197 0.221 0.238 0.224 0.268 0.294 0.268 0.218 0.221 0.253 0.250 0.279 0.253 0.253 0.238 0.232 0.235 0.238 0.253 0.247 0.235 0.268 0.259 0.274 0.262 0.279 0.247 0.226 0.276 0.279 0.022 0.024 0.024 0.025 0.023 0.024 0.024 0.022 0.025 0.025 0.024 0.025 0.023 0.022 EA0270R Lag B2 Ophthalmia Range PC033 0.259 0.238 0.232 0.232 0.235 0.247 0.262 0.204 0.259 0.292 0.223 0.241 0.268 0.214 0.259 0.250 0.268 0.256 0.274 0.253 0.241 0.226 0.253 0.256 0.208 0.235 0.259 0.244 0.241 0.235 0.229 0.235 0.265 0.226 0.277 0.208 0.211 0.214 0.205 0.238 0.265 0.202 0.202 0.217 0.262 0.274 0.023 0.025 0.023 0.019 0.024 0.022 0.021 0.022 0.021 0.022 0.026 0.024 0.021 YYHC0048I G87 LN8425 PC034 0.231 0.204 0.229 0.213 0.211 0.231 0.211 0.189 0.201 0.215 0.199 0.237 0.248 0.230 0.235 0.230 0.219 0.208 0.217 0.226 0.208 0.216 0.201 0.204 0.186 0.213 0.213 0.224 0.215 0.129 0.129 0.129 0.114 0.138 0.261 0.208 0.215 0.206 0.195 0.140 0.232 0.202 0.218 0.233 0.244 0.259 0.244 0.027 0.023 0.023 0.022 0.020 0.023 0.022 0.017 0.019 0.020 0.017 0.019 MG0165R J8 31 Tyrannochthonius sp PC035 0.285 0.271 0.241 0.253 0.241 0.232 0.300 0.252 0.309 0.306 0.241 0.235 0.276 0.247 0.268 0.265 0.274 0.274 0.262 0.268 0.265 0.309 0.279 0.279 0.259 0.238 0.276 0.244 0.238 0.268 0.262 0.271 0.262 0.274 0.303 0.256 0.259 0.271 0.276 0.268 0.271 0.250 0.268 0.265 0.282 0.280 0.301 0.309 0.023 0.025 0.023 0.024 0.024 0.025 0.026 0.022 0.025 0.026 0.022 EJR0202 2 9 9 Tyrranochthonius B2 PC036 0.252 0.214 0.217 0.214 0.214 0.220 0.243 0.237 0.229 0.255 0.217 0.258 0.240 0.249 0.287 0.290 0.290 0.284 0.276 0.238 0.214 0.261 0.252 0.255 0.217 0.217 0.196 0.226 0.232 0.246 0.249 0.246 0.243 0.235 0.264 0.214 0.214 0.217 0.208 0.226 0.223 0.232 0.226 0.205 0.243 0.291 0.259 0.249 0.250 0.023 0.022 0.024 0.023 0.019 0.024 0.024 0.026 0.022 0.022 LB084 Tyran Wheelarra Hill PC037 0.207 0.201 0.207 0.186 0.201 0.213 0.237 0.182 0.210 0.269 0.201 0.234 0.249 0.216 0.246 0.234 0.249 0.254 0.254 0.204 0.219 0.219 0.207 0.225 0.198 0.186 0.169 0.180 0.166 0.207 0.210 0.198 0.204 0.207 0.237 0.201 0.201 0.213 0.195 0.195 0.213 0.198 0.160 0.180 0.222 0.246 0.195 0.222 0.258 0.163 0.018 0.021 0.022 0.022 0.023 0.019 0.025 0.022 0.021 PI012 Tyran Wheelarra Hill PC038 0.213 0.210 0.207 0.219 0.195 0.207 0.222 0.152 0.231 0.257 0.192 0.237 0.222 0.186 0.234 0.243 0.260 0.246 0.263 0.228 0.178 0.210 0.216 0.246 0.183 0.207 0.204 0.204 0.189 0.207 0.204 0.213 0.210 0.213 0.216 0.189 0.192 0.195 0.189 0.201 0.210 0.216 0.160 0.195 0.231 0.267 0.228 0.216 0.237 0.195 0.154 0.020 0.021 0.020 0.024 0.018 0.025 0.020 0.022 EXR1148 J20-6 Lagy S4 Hancock Range North PC039 0.234 0.222 0.237 0.228 0.204 0.231 0.216 0.206 0.219 0.240 0.201 0.249 0.254 0.231 0.234 0.234 0.240 0.243 0.240 0.225 0.222 0.189 0.228 0.228 0.213 0.207 0.198 0.219 0.210 0.183 0.201 0.204 0.213 0.210 0.216 0.213 0.222 0.219 0.225 0.222 0.243 0.198 0.178 0.219 0.216 0.264 0.225 0.216 0.291 0.231 0.197 0.197 0.023 0.019 0.020 0.021 0.024 0.021 0.023 GF0173R Lag B2 Jirrpalpur Range PC040 0.229 0.185 0.243 0.235 0.214 0.246 0.223 0.210 0.235 0.279 0.194 0.235 0.243 0.232 0.258 0.249 0.267 0.273 0.264 0.238 0.194 0.188 0.255 0.249 0.208 0.235 0.223 0.249 0.240 0.211 0.223 0.226 0.229 0.238 0.235 0.205 0.211 0.220 0.208 0.229 0.223 0.188 0.155 0.205 0.261 0.265 0.238 0.252 0.288 0.226 0.204 0.183 0.189 0.019 0.021 0.022 0.025 0.023 0.021 JIN0078R BHP772 Lagynochthonius Hancock Range South PC041 0.267 0.211 0.235 0.229 0.217 0.246 0.267 0.207 0.238 0.276 0.191 0.226 0.261 0.243 0.287 0.282 0.305 0.284 0.284 0.243 0.196 0.208 0.261 0.252 0.185 0.246 0.214 0.246 0.243 0.220 0.226 0.226 0.264 0.232 0.282 0.223 0.232 0.220 0.220 0.243 0.240 0.202 0.191 0.226 0.282 0.315 0.232 0.246 0.300 0.214 0.198 0.180 0.195 0.147 0.020 0.021 0.025 0.022 0.023 AX83 10DDH033P1T1-2 PC042 0.202 0.203 0.205 0.200 0.196 0.237 0.245 0.199 0.259 0.224 0.217 0.242 0.264 0.189 0.212 0.225 0.229 0.216 0.220 0.223 0.189 0.187 0.194 0.194 0.189 0.212 0.228 0.190 0.191 0.190 0.199 0.212 0.199 0.209 0.243 0.186 0.248 0.207 0.200 0.204 0.242 0.204 0.199 0.203 0.223 0.250 0.220 0.191 0.271 0.240 0.237 0.201 0.219 0.246 0.232 0.014 0.015 0.014 0.020 AX82 ID31P1T1-5 PC043 0.194 0.201 0.216 0.200 0.207 0.190 0.223 0.157 0.216 0.230 0.174 0.240 0.244 0.195 0.200 0.195 0.209 0.217 0.215 0.186 0.177 0.170 0.202 0.206 0.186 0.202 0.220 0.198 0.200 0.175 0.173 0.175 0.175 0.193 0.263 0.174 0.201 0.199 0.194 0.181 0.204 0.189 0.177 0.206 0.208 0.243 0.225 0.208 0.279 0.251 0.179 0.176 0.188 0.207 0.213 0.201 0.015 0.014 0.018 AX86 ID32P1T1-7 PC044 0.211 0.226 0.235 0.222 0.218 0.236 0.191 0.220 0.247 0.244 0.226 0.262 0.261 0.220 0.224 0.213 0.209 0.240 0.229 0.231 0.197 0.186 0.213 0.224 0.209 0.233 0.248 0.228 0.235 0.209 0.209 0.222 0.202 0.213 0.263 0.222 0.213 0.204 0.211 0.210 0.244 0.219 0.200 0.218 0.235 0.285 0.249 0.210 0.309 0.287 0.239 0.224 0.223 0.246 0.257 0.221 0.145 0.017 0.020 AX85 ID32P1T1-7 PC045 0.187 0.185 0.187 0.177 0.179 0.211 0.196 0.140 0.212 0.226 0.176 0.231 0.229 0.191 0.238 0.238 0.243 0.220 0.214 0.174 0.158 0.176 0.220 0.214 0.169 0.189 0.202 0.178 0.172 0.183 0.194 0.189 0.183 0.196 0.240 0.168 0.205 0.193 0.184 0.179 0.230 0.178 0.176 0.169 0.199 0.256 0.232 0.188 0.265 0.217 0.186 0.183 0.186 0.223 0.229 0.180 0.170 0.204 0.016 JV08 DD15BS40010-20160608-T2-02 PC046 0.218 0.185 0.194 0.187 0.189 0.244 0.245 0.212 0.256 0.242 0.236 0.228 0.255 0.172 0.162 0.164 0.167 0.164 0.162 0.218 0.200 0.213 0.187 0.184 0.189 0.204 0.251 0.184 0.173 0.192 0.203 0.205 0.205 0.229 0.284 0.168 0.182 0.167 0.187 0.206 0.227 0.219 0.201 0.238 0.216 0.250 0.256 0.213 0.262 0.252 0.246 0.237 0.243 0.264 0.279 0.194 0.197 0.235 0.205 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 7. Estimates of Evolutionary Divergence between Sequences

Lineage 127103 IV221 IV265 IV224 IV259 IV216 IV222 AX81 DF18 Isp FZ1 FZ2 G107 G211 GB0018R GC0103R IT72 IV201 IV202 IV206 IV209 IV211 IV213 IV217 IV219 JD01 JF56 PSF0090R 127103 Pseudoscorpiones gp 9 0.017 0.016 0.019 0.016 0.017 0.016 0.016 0.021 0.021 0.021 0.020 0.024 0.021 0.025 0.026 0.020 0.019 0.017 0.020 0.019 0.016 0.018 0.019 0.018 0.020 0.019 0.024 IV221 139896 Pseudoscorpiones gp 10 0.180 0.010 0.013 0.011 0.013 0.010 0.018 0.021 0.020 0.021 0.021 0.024 0.021 0.027 0.027 0.017 0.020 0.019 0.021 0.020 0.016 0.015 0.020 0.012 0.020 0.014 0.027 IV265 RC15MEB0020-20160317-T2-02 Pseudoscorpiones gp 11 0.206 0.053 0.013 0.010 0.011 0.010 0.016 0.021 0.019 0.019 0.019 0.023 0.020 0.026 0.025 0.018 0.018 0.018 0.020 0.020 0.014 0.017 0.018 0.012 0.017 0.016 0.025 IV224 138538 Pseudoscorpiones gp 12 0.214 0.055 0.079 0.009 0.011 0.008 0.017 0.026 0.022 0.025 0.024 0.030 0.025 0.028 0.028 0.020 0.025 0.025 0.024 0.024 0.019 0.019 0.026 0.014 0.023 0.019 0.029 IV259 DD15MEB0018-20160317-T3-01 Pseudoscorpiones gp 12-2 0.206 0.065 0.070 0.036 0.009 0.007 0.015 0.022 0.020 0.021 0.021 0.025 0.022 0.025 0.025 0.017 0.019 0.021 0.020 0.020 0.015 0.015 0.020 0.009 0.019 0.016 0.024 IV216 138532 Pseudoscorpiones gp 13 0.216 0.076 0.095 0.075 0.069 0.007 0.016 0.021 0.019 0.022 0.022 0.024 0.021 0.025 0.025 0.017 0.020 0.020 0.021 0.020 0.017 0.016 0.020 0.011 0.020 0.015 0.026 IV222 138537 Pseudoscorpiones gp 14 0.207 0.048 0.072 0.042 0.044 0.035 0.015 0.022 0.019 0.021 0.021 0.024 0.021 0.026 0.026 0.015 0.019 0.020 0.021 0.021 0.016 0.015 0.020 0.009 0.019 0.015 0.026 AX81 ID27P1T4-5 Pseudoscorpionida 0.236 0.203 0.222 0.206 0.217 0.223 0.204 0.021 0.018 0.019 0.019 0.024 0.018 0.026 0.025 0.019 0.018 0.021 0.019 0.019 0.018 0.021 0.019 0.018 0.018 0.020 0.025 DF18 8668 Indohya sp. 0.220 0.228 0.225 0.225 0.236 0.243 0.234 0.207 0.020 0.020 0.020 0.019 0.022 0.023 0.024 0.022 0.018 0.022 0.018 0.018 0.020 0.020 0.019 0.021 0.019 0.021 0.024 EU559564 Indohya sp. JM-2008 0.240 0.219 0.224 0.204 0.224 0.217 0.203 0.187 0.215 0.020 0.020 0.021 0.020 0.025 0.025 0.019 0.019 0.020 0.020 0.019 0.020 0.019 0.017 0.018 0.019 0.020 0.024 FZ1 Pseudoscorpion RC13KOOD0302P7T1-1 0.264 0.214 0.214 0.222 0.228 0.228 0.209 0.221 0.200 0.228 0.005 0.021 0.021 0.023 0.022 0.023 0.019 0.020 0.020 0.019 0.019 0.020 0.020 0.020 0.011 0.020 0.024 FZ2 Pseudoscorpion RC13KOOD0305P7T3-3 0.264 0.214 0.214 0.222 0.228 0.228 0.209 0.221 0.204 0.228 0.010 0.022 0.021 0.023 0.023 0.023 0.019 0.020 0.019 0.019 0.018 0.020 0.021 0.019 0.012 0.020 0.024 G107 Indohya 0.267 0.279 0.279 0.275 0.276 0.293 0.282 0.287 0.199 0.252 0.258 0.261 0.020 0.024 0.023 0.025 0.021 0.024 0.018 0.017 0.024 0.023 0.022 0.024 0.020 0.022 0.023 G211 Pseudoscorpian 109256 0.256 0.228 0.225 0.232 0.237 0.251 0.232 0.199 0.230 0.244 0.225 0.228 0.255 0.023 0.022 0.023 0.020 0.020 0.019 0.020 0.022 0.022 0.019 0.021 0.019 0.022 0.022 GB0018R Hyidae B1 0.349 0.308 0.296 0.288 0.308 0.308 0.308 0.287 0.255 0.293 0.238 0.240 0.276 0.282 0.012 0.024 0.023 0.027 0.024 0.023 0.024 0.023 0.022 0.026 0.021 0.025 0.021 GC0103R Hyidae 0.323 0.293 0.284 0.280 0.302 0.308 0.305 0.267 0.258 0.276 0.235 0.238 0.258 0.249 0.053 0.024 0.023 0.025 0.022 0.023 0.024 0.024 0.022 0.026 0.021 0.024 0.020 IT72 BHRC0512-151013-T3-01 Pseudoscorpiones 0.194 0.109 0.110 0.093 0.113 0.116 0.102 0.196 0.215 0.192 0.230 0.227 0.268 0.235 0.287 0.277 0.022 0.020 0.022 0.022 0.017 0.016 0.021 0.015 0.020 0.016 0.027 IV201 DD14MEB0002-20150607-01sc Pseudoscorpiones 0.254 0.235 0.236 0.222 0.236 0.238 0.223 0.207 0.216 0.215 0.224 0.228 0.258 0.242 0.252 0.255 0.245 0.019 0.018 0.018 0.020 0.020 0.010 0.020 0.018 0.019 0.021 IV202 DD14MRR0004T1-04 Pseudoscorpiones 0.163 0.240 0.229 0.215 0.225 0.238 0.234 0.245 0.236 0.252 0.264 0.269 0.284 0.258 0.337 0.326 0.231 0.267 0.020 0.021 0.018 0.018 0.019 0.019 0.020 0.018 0.024 IV206 DD14MRR0008-20150808-T1-02 Pseudoscorpiones 0.234 0.251 0.256 0.222 0.256 0.258 0.249 0.209 0.162 0.219 0.231 0.236 0.176 0.227 0.264 0.246 0.263 0.213 0.262 0.009 0.022 0.021 0.019 0.021 0.017 0.021 0.023 IV209 M2ERC027-t2-03 Pseudoscorpiones 0.227 0.244 0.249 0.218 0.245 0.258 0.245 0.207 0.162 0.215 0.228 0.228 0.167 0.237 0.261 0.249 0.263 0.207 0.262 0.036 0.021 0.021 0.019 0.021 0.018 0.020 0.022 IV211 RC13MEA0279-20150807-T1-02 Pseudoscorpiones 0.185 0.118 0.122 0.126 0.140 0.134 0.125 0.200 0.218 0.219 0.221 0.216 0.284 0.249 0.290 0.293 0.113 0.227 0.224 0.240 0.227 0.015 0.018 0.016 0.019 0.014 0.026 IV213 DD14MRR0009-20151002-T2-02 Pseudoscorpiones 0.212 0.122 0.125 0.116 0.143 0.138 0.125 0.207 0.220 0.233 0.228 0.224 0.308 0.232 0.305 0.302 0.105 0.253 0.227 0.249 0.240 0.131 0.020 0.015 0.019 0.011 0.026 IV217 RC14MEB0029-20151001-T1-01 Pseudoscorpiones 0.256 0.244 0.236 0.239 0.241 0.243 0.227 0.212 0.218 0.215 0.233 0.240 0.255 0.246 0.255 0.255 0.247 0.049 0.260 0.227 0.222 0.224 0.258 0.020 0.018 0.019 0.023 IV219 RC14MEB0101-20151001-T1-02 Pseudoscorpiones 0.186 0.066 0.080 0.048 0.045 0.066 0.042 0.212 0.254 0.220 0.215 0.210 0.282 0.232 0.305 0.296 0.113 0.252 0.242 0.261 0.252 0.141 0.134 0.254 0.019 0.015 0.025 JD01 RC13KOOD0184-151011-T1-03 Pseudoscorpion 0.249 0.210 0.198 0.212 0.209 0.218 0.198 0.192 0.189 0.201 0.060 0.065 0.246 0.215 0.229 0.211 0.204 0.196 0.247 0.211 0.204 0.204 0.209 0.198 0.209 0.018 0.022 JF56 RC14MEH0252-20160120-T1-04 Psuedoscorpiones 0.212 0.122 0.122 0.123 0.140 0.136 0.131 0.200 0.227 0.226 0.243 0.238 0.299 0.244 0.299 0.293 0.099 0.249 0.216 0.253 0.251 0.129 0.069 0.249 0.141 0.202 0.025 PSF0090R LN2124 Packsaddle Indohya 0.343 0.331 0.305 0.322 0.320 0.317 0.317 0.305 0.282 0.287 0.276 0.273 0.287 0.258 0.173 0.176 0.283 0.273 0.349 0.258 0.258 0.311 0.299 0.279 0.323 0.252 0.314 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 12. Estimates of Evolutionary Divergence between Sequences

Specimen ID IV289 IV295 IV293 IV326 WAMT137066 IV61 IV07 IV82 IV76 IV09 IV79 IV17 IV14 IV27 IV55 IV81 Dbram Dmeso Pana Pbyth Pgnop Pkryp Psp C119 C37 C58 FT4 FT5 FT6 FZ21 GG9 GH7 GH8 HE17 HE4 IS15 IS33 IT67 Pcard U72 U74 U83 IV289 110827 Schizomid gp 1 Paradraculoides bythius 0.011 0.011 0.013 0.016 0.003 0.009 0.010 0.011 0.012 0.013 0.013 0.012 0.013 0.012 0.012 0.013 0.013 0.011 0.003 0.011 0.010 0.010 0.011 0.012 0.011 0.012 0.014 0.012 0.013 0.013 0.013 0.012 0.011 0.011 0.011 0.012 0.011 0.012 0.013 0.013 0.012 IV295 133446 Schizomid gp 2 Paradraculoides anachoretus 0.110 0.008 0.013 0.017 0.011 0.010 0.010 0.011 0.012 0.013 0.012 0.011 0.012 0.013 0.013 0.012 0.012 0.003 0.011 0.012 0.012 0.010 0.012 0.012 0.012 0.011 0.012 0.011 0.012 0.012 0.011 0.012 0.012 0.012 0.013 0.011 0.011 0.013 0.012 0.012 0.012 IV293 140982 Schizomid gp 3 P. sp.'SCH034' 0.085 0.056 0.014 0.017 0.010 0.010 0.010 0.011 0.011 0.014 0.012 0.012 0.012 0.013 0.012 0.013 0.013 0.008 0.010 0.011 0.011 0.010 0.012 0.012 0.012 0.012 0.013 0.012 0.013 0.012 0.011 0.012 0.012 0.013 0.013 0.012 0.012 0.014 0.012 0.012 0.012 IV326 DD15MEB0008-20160317-T2-01 Schizomid gp 4 Bamazomus sp. 0.169 0.163 0.160 0.016 0.013 0.013 0.013 0.013 0.013 0.000 0.012 0.014 0.012 0.013 0.013 0.012 0.013 0.014 0.014 0.013 0.014 0.013 0.014 0.013 0.014 0.013 0.013 0.012 0.014 0.012 0.012 0.013 0.012 0.013 0.013 0.013 0.013 0.014 0.012 0.012 0.012 WAMT137066 Schizomid gp 5 B. subsolanus 0.205 0.205 0.205 0.179 0.016 0.018 0.016 0.015 0.015 0.016 0.014 0.015 0.016 0.016 0.016 0.015 0.016 0.017 0.017 0.016 0.016 0.016 0.017 0.016 0.016 0.017 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.016 0.017 0.016 0.014 0.015 0.015 0.015 IV61 DD14MEC0002-20150929-T3-03 lin 1 0.008 0.108 0.086 0.161 0.201 0.009 0.009 0.011 0.011 0.013 0.013 0.012 0.013 0.013 0.013 0.013 0.014 0.011 0.001 0.011 0.010 0.010 0.012 0.013 0.011 0.012 0.013 0.012 0.013 0.012 0.012 0.012 0.011 0.011 0.011 0.011 0.011 0.012 0.013 0.013 0.012 IV07 Budgie20150604-06 lin 2 0.070 0.078 0.074 0.156 0.210 0.069 0.009 0.012 0.011 0.013 0.013 0.012 0.012 0.013 0.012 0.013 0.013 0.010 0.009 0.011 0.011 0.010 0.012 0.012 0.011 0.013 0.014 0.012 0.014 0.013 0.012 0.012 0.012 0.012 0.012 0.012 0.012 0.013 0.013 0.013 0.013 IV82 TOBRC0023-20151001-T1-01 lin 3 0.086 0.088 0.080 0.160 0.205 0.081 0.063 0.010 0.011 0.013 0.014 0.012 0.013 0.012 0.013 0.013 0.013 0.011 0.010 0.010 0.011 0.007 0.012 0.012 0.011 0.012 0.013 0.012 0.013 0.012 0.012 0.012 0.011 0.012 0.012 0.012 0.012 0.014 0.012 0.011 0.013 IV76 MELUNK11-20151001-T2-01 lin 4 0.103 0.119 0.105 0.160 0.190 0.102 0.111 0.107 0.007 0.013 0.013 0.012 0.013 0.012 0.014 0.012 0.013 0.011 0.011 0.010 0.011 0.011 0.013 0.011 0.012 0.013 0.013 0.013 0.013 0.013 0.012 0.012 0.011 0.013 0.011 0.012 0.012 0.014 0.012 0.012 0.012 IV09 DD14MEL0001-t2-01 lin 4a 0.122 0.116 0.102 0.166 0.186 0.117 0.108 0.110 0.034 0.013 0.013 0.013 0.013 0.013 0.013 0.012 0.013 0.012 0.012 0.010 0.010 0.011 0.013 0.011 0.011 0.013 0.013 0.013 0.012 0.013 0.012 0.012 0.011 0.013 0.012 0.012 0.012 0.014 0.012 0.012 0.012 IV79 RC14MEB0060-20151001-T2-01 lin 5 0.169 0.163 0.160 0.000 0.179 0.161 0.156 0.160 0.160 0.166 0.012 0.014 0.012 0.013 0.013 0.012 0.013 0.014 0.014 0.013 0.014 0.013 0.014 0.013 0.014 0.013 0.013 0.012 0.014 0.012 0.012 0.013 0.012 0.013 0.013 0.013 0.013 0.014 0.012 0.012 0.012 IV17 DD14MRR007-20150808-T1-02 lin 6 0.167 0.149 0.147 0.172 0.194 0.166 0.156 0.152 0.152 0.158 0.172 0.011 0.010 0.012 0.013 0.012 0.011 0.013 0.013 0.013 0.013 0.014 0.014 0.012 0.012 0.013 0.013 0.012 0.012 0.013 0.012 0.012 0.013 0.012 0.013 0.012 0.013 0.013 0.011 0.011 0.012 IV14 DD14MRR0005-T1-01 lin 7 0.174 0.160 0.152 0.178 0.201 0.172 0.166 0.166 0.146 0.153 0.178 0.091 0.009 0.012 0.013 0.012 0.009 0.012 0.012 0.014 0.012 0.012 0.013 0.012 0.013 0.014 0.014 0.013 0.012 0.014 0.013 0.013 0.013 0.012 0.012 0.012 0.013 0.013 0.012 0.012 0.012 IV27 M2ERC0053-T2-02 lin 8 0.167 0.146 0.139 0.160 0.220 0.167 0.152 0.153 0.139 0.147 0.160 0.086 0.072 0.012 0.014 0.012 0.003 0.012 0.013 0.013 0.013 0.014 0.014 0.012 0.013 0.014 0.013 0.013 0.012 0.013 0.013 0.013 0.013 0.012 0.013 0.012 0.012 0.012 0.012 0.012 0.012 IV55 RC13MRR0077-T1-02 lin 9 0.164 0.160 0.156 0.183 0.190 0.163 0.152 0.150 0.129 0.131 0.183 0.122 0.124 0.119 0.013 0.012 0.012 0.013 0.013 0.013 0.012 0.013 0.013 0.012 0.013 0.014 0.014 0.014 0.014 0.014 0.013 0.012 0.013 0.012 0.012 0.012 0.013 0.014 0.013 0.013 0.014 IV81 RC14MRR0017-20151002-T2-01 lin 10 0.158 0.152 0.152 0.182 0.201 0.156 0.153 0.155 0.160 0.167 0.182 0.150 0.172 0.161 0.172 0.013 0.014 0.013 0.013 0.012 0.013 0.013 0.013 0.013 0.013 0.012 0.012 0.012 0.013 0.012 0.013 0.013 0.013 0.013 0.013 0.012 0.013 0.015 0.013 0.013 0.013 EU272684 Draculoides bramstokeri 0.157 0.141 0.138 0.155 0.184 0.152 0.148 0.154 0.132 0.135 0.155 0.146 0.148 0.144 0.152 0.165 0.012 0.012 0.013 0.012 0.012 0.012 0.013 0.012 0.012 0.013 0.013 0.013 0.012 0.013 0.013 0.013 0.011 0.012 0.012 0.011 0.012 0.012 0.012 0.012 0.013 EU272730 Draculoides mesozeirus SzJ 0.165 0.146 0.139 0.157 0.217 0.165 0.149 0.151 0.136 0.141 0.157 0.089 0.073 0.006 0.120 0.162 0.141 0.012 0.013 0.013 0.013 0.014 0.014 0.012 0.014 0.014 0.014 0.013 0.013 0.014 0.013 0.013 0.012 0.012 0.014 0.012 0.012 0.013 0.012 0.013 0.012 Panachoretus WAM T66236 Paradraculoides 0.115 0.005 0.063 0.169 0.209 0.113 0.085 0.095 0.124 0.121 0.169 0.155 0.165 0.152 0.166 0.160 0.147 0.151 0.011 0.012 0.012 0.010 0.012 0.013 0.013 0.012 0.013 0.012 0.012 0.012 0.012 0.012 0.012 0.012 0.013 0.011 0.012 0.014 0.013 0.013 0.012 Pbythius WAM T63344 Paradraculoides byth 0.006 0.110 0.087 0.165 0.206 0.002 0.069 0.085 0.103 0.119 0.165 0.169 0.171 0.168 0.166 0.161 0.156 0.164 0.111 0.011 0.010 0.010 0.012 0.013 0.011 0.013 0.014 0.012 0.014 0.013 0.013 0.012 0.012 0.012 0.011 0.012 0.011 0.012 0.014 0.013 0.013 Pgnophicola WAM T63371 3 Paradraculoides 0.105 0.116 0.102 0.174 0.202 0.100 0.087 0.076 0.090 0.095 0.174 0.135 0.161 0.148 0.131 0.150 0.137 0.144 0.121 0.102 0.010 0.010 0.012 0.012 0.011 0.012 0.013 0.012 0.012 0.012 0.012 0.012 0.012 0.012 0.012 0.011 0.013 0.012 0.013 0.013 0.014 Pkryptus WAM T65802 Paradraculoides kryp 0.111 0.113 0.087 0.174 0.193 0.110 0.102 0.111 0.095 0.100 0.174 0.152 0.150 0.153 0.142 0.165 0.140 0.152 0.118 0.108 0.092 0.010 0.013 0.011 0.012 0.014 0.013 0.013 0.013 0.013 0.012 0.012 0.012 0.013 0.012 0.012 0.012 0.013 0.012 0.012 0.013 Psp OFB 2007 Paradraculoides sp. OFB-200 0.094 0.089 0.084 0.153 0.197 0.089 0.074 0.039 0.097 0.103 0.153 0.153 0.166 0.160 0.150 0.156 0.137 0.156 0.094 0.090 0.084 0.098 0.013 0.012 0.012 0.013 0.014 0.013 0.012 0.013 0.013 0.014 0.012 0.013 0.011 0.012 0.012 0.012 0.013 0.013 0.013 C119 ex T95543 0.132 0.144 0.141 0.173 0.212 0.137 0.130 0.146 0.130 0.143 0.173 0.167 0.170 0.165 0.171 0.144 0.162 0.162 0.150 0.137 0.134 0.148 0.147 0.012 0.013 0.012 0.013 0.013 0.013 0.012 0.012 0.011 0.013 0.012 0.013 0.012 0.012 0.013 0.012 0.012 0.012 C37 T92541B 0.146 0.139 0.133 0.186 0.197 0.144 0.127 0.133 0.129 0.127 0.186 0.151 0.164 0.153 0.144 0.153 0.155 0.152 0.142 0.140 0.116 0.123 0.119 0.156 0.012 0.013 0.013 0.013 0.013 0.013 0.012 0.012 0.012 0.012 0.012 0.010 0.013 0.012 0.011 0.012 0.012 C58 T98320 0.108 0.142 0.127 0.172 0.173 0.105 0.119 0.119 0.132 0.130 0.172 0.160 0.167 0.163 0.150 0.161 0.148 0.157 0.150 0.108 0.118 0.123 0.121 0.141 0.120 0.013 0.014 0.013 0.014 0.013 0.013 0.013 0.012 0.012 0.012 0.012 0.012 0.013 0.012 0.011 0.011 FT4 Draculoides SCH012 PSB0166R 0.167 0.141 0.142 0.161 0.214 0.169 0.149 0.153 0.160 0.160 0.161 0.150 0.171 0.160 0.169 0.133 0.143 0.162 0.148 0.174 0.155 0.171 0.150 0.135 0.175 0.166 0.008 0.006 0.012 0.007 0.013 0.013 0.013 0.012 0.012 0.012 0.012 0.015 0.014 0.014 0.013 FT5 Draculoides sp PSB0163R 0.164 0.152 0.152 0.166 0.207 0.163 0.146 0.146 0.146 0.149 0.166 0.161 0.167 0.161 0.160 0.136 0.140 0.159 0.160 0.168 0.139 0.152 0.148 0.132 0.173 0.175 0.064 0.008 0.013 0.009 0.013 0.012 0.014 0.012 0.013 0.013 0.013 0.014 0.013 0.013 0.014 FT6 Draculoides SCH012 GBP553 0.164 0.139 0.136 0.147 0.203 0.163 0.146 0.147 0.146 0.147 0.147 0.161 0.164 0.158 0.161 0.138 0.143 0.159 0.147 0.168 0.155 0.160 0.144 0.132 0.178 0.175 0.030 0.063 0.012 0.006 0.013 0.013 0.013 0.013 0.012 0.013 0.013 0.014 0.013 0.014 0.013 FZ21 Schizomida RC13KOOD0308P7T1-1 0.155 0.153 0.141 0.144 0.201 0.150 0.142 0.144 0.149 0.158 0.144 0.170 0.182 0.152 0.185 0.144 0.138 0.152 0.160 0.155 0.145 0.152 0.139 0.149 0.170 0.181 0.122 0.117 0.118 0.012 0.013 0.013 0.013 0.014 0.013 0.013 0.013 0.015 0.014 0.014 0.013 GG9 Draculoides sp PSA0806R 0.169 0.136 0.142 0.150 0.205 0.167 0.147 0.146 0.149 0.155 0.150 0.144 0.164 0.156 0.160 0.133 0.135 0.159 0.144 0.173 0.152 0.161 0.142 0.137 0.167 0.171 0.031 0.061 0.028 0.117 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.015 0.013 0.014 0.013 GH7 Draculoides Dra-PES16433 0.146 0.138 0.136 0.169 0.192 0.146 0.131 0.136 0.139 0.139 0.169 0.172 0.175 0.164 0.164 0.146 0.162 0.160 0.140 0.148 0.152 0.142 0.145 0.135 0.138 0.133 0.153 0.147 0.147 0.156 0.147 0.007 0.012 0.013 0.012 0.013 0.012 0.013 0.013 0.013 0.012 GH8 Draculoides Dra-PES16447 0.142 0.128 0.125 0.161 0.194 0.146 0.121 0.135 0.130 0.136 0.161 0.163 0.155 0.142 0.152 0.150 0.160 0.139 0.132 0.147 0.139 0.132 0.145 0.119 0.141 0.130 0.146 0.133 0.136 0.147 0.146 0.038 0.012 0.012 0.013 0.013 0.012 0.013 0.013 0.013 0.012 HE17 BHRC151-20141216-T3-01c 0.129 0.135 0.137 0.175 0.179 0.126 0.129 0.123 0.127 0.124 0.175 0.143 0.157 0.148 0.151 0.148 0.149 0.146 0.141 0.128 0.123 0.111 0.121 0.148 0.115 0.118 0.164 0.156 0.156 0.156 0.159 0.137 0.137 0.012 0.012 0.011 0.012 0.013 0.012 0.012 0.013 HE4 BHRC001-20141028-SC01 0.137 0.143 0.143 0.165 0.207 0.137 0.138 0.129 0.134 0.143 0.165 0.142 0.149 0.138 0.153 0.146 0.165 0.130 0.147 0.137 0.131 0.132 0.132 0.144 0.121 0.118 0.165 0.143 0.160 0.162 0.154 0.145 0.145 0.115 0.011 0.011 0.011 0.013 0.011 0.011 0.012 IS15 KBRC1389 20150909 04 Schizomida 0.131 0.135 0.131 0.155 0.207 0.124 0.122 0.124 0.122 0.127 0.155 0.150 0.161 0.135 0.161 0.149 0.133 0.136 0.142 0.127 0.121 0.127 0.126 0.138 0.135 0.109 0.147 0.166 0.150 0.156 0.144 0.152 0.144 0.124 0.124 0.011 0.011 0.012 0.013 0.013 0.013 IS33 Kbrc116720150809t2 05 Schizomida sp 0.122 0.128 0.125 0.156 0.186 0.121 0.124 0.119 0.121 0.130 0.156 0.139 0.155 0.139 0.152 0.142 0.130 0.139 0.129 0.118 0.111 0.111 0.121 0.124 0.114 0.108 0.146 0.153 0.149 0.159 0.144 0.138 0.122 0.104 0.099 0.100 0.011 0.012 0.011 0.011 0.012 IT67 BHRC401-151013-T1-01 Schizomida 0.121 0.114 0.125 0.157 0.192 0.116 0.114 0.116 0.111 0.121 0.157 0.146 0.157 0.144 0.149 0.147 0.135 0.141 0.121 0.119 0.116 0.108 0.111 0.130 0.132 0.113 0.155 0.138 0.154 0.144 0.150 0.139 0.127 0.115 0.110 0.099 0.105 0.013 0.012 0.011 0.012 T93233 C17 CBRC099P5T1-3 Paradraculoides sp cardo 0.132 0.119 0.119 0.148 0.162 0.123 0.117 0.125 0.134 0.126 0.148 0.130 0.144 0.152 0.143 0.170 0.146 0.150 0.119 0.123 0.115 0.112 0.114 0.161 0.130 0.106 0.166 0.168 0.155 0.162 0.162 0.139 0.125 0.121 0.135 0.116 0.114 0.105 0.014 0.014 0.015 U72 Shizomid T95463 0.147 0.131 0.128 0.161 0.190 0.146 0.124 0.125 0.132 0.131 0.161 0.121 0.146 0.122 0.133 0.147 0.124 0.120 0.139 0.152 0.129 0.152 0.131 0.129 0.142 0.138 0.130 0.133 0.133 0.147 0.124 0.155 0.149 0.138 0.140 0.133 0.130 0.129 0.134 0.004 0.010 U74 Shizomid T95462 0.136 0.139 0.128 0.163 0.197 0.135 0.127 0.127 0.129 0.130 0.163 0.133 0.152 0.124 0.139 0.158 0.133 0.122 0.147 0.140 0.135 0.152 0.132 0.129 0.145 0.130 0.139 0.140 0.140 0.150 0.130 0.152 0.150 0.146 0.142 0.130 0.132 0.125 0.137 0.018 0.010 U83 Shizomid T95450 0.147 0.125 0.125 0.160 0.194 0.149 0.124 0.125 0.130 0.133 0.160 0.144 0.153 0.131 0.147 0.144 0.137 0.125 0.131 0.150 0.140 0.152 0.132 0.137 0.153 0.138 0.127 0.141 0.137 0.142 0.122 0.147 0.146 0.151 0.154 0.145 0.141 0.124 0.141 0.078 0.081 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Molecular systematics of subfauna from the Robe Valley

1 October, 2016

Jason Alexander Biota Environmental Sciences Level 1, 228 Carr Place Leederville, WA 6007

Via email

Re. Report on the molecular systematics of subfauna from the Robe River Valley – historical, Phase 3 and Phase 1 and 2 repeats: Part 2

Dear Jason,

Following is a summary of the results of the subfauna study we have completed on three taxonomic groups from the Robe River Valley (Symphyla, Chilopoda, Coleoptera). Seventeen distinct genetic lineages were detected among the three groups. The 17 lineages likely represent 17 species, four or five of which have been detected previously in the Pilbara.

Sincerely,

Dr. Terrie Finston, Yvette Hitchen and Dr. Oliver Berry Helix Molecular Solutions

1 Molecular systematics of subfauna from the Robe Valley

Background and Objective Sixty-two specimens of subfauna belonging to three taxonomic groups (Chilopoda, Coleoptera, Symphyla) were collected from Robe River Valley and sequenced for variation at the mitochondrial COXI gene. The molecular data were assessed in order to determine the number of species present in each group and compare the results to those obtained during previous surveys that have been undertaken for the these groups elsewhere in the Pilbara.

Executive summary  Sixty-two specimens from the Robe River Valley belonging to three taxonomic groups were sequenced for variation at the COI gene.  Seventeen lineages were detected among the three groups.  The 17 lineages likely represent 17 species.  Four or five of the species have been detected previously in the Pilbara, whereas the remainder appear to be new, based on the material available for comparison.

Results Symphyla Preliminary analysis - Reference sequences and outgroups Nine specimens of Symphyla from the Robe Valley (historical, phase 3, phase 1 and 2 repeats) were sequenced for COI (Table 18). Two specimens yielded a DNA sequence, and a search of similar sequences on Genbank and in the Helix database indicated that one was most similar to the family Scutigerellidae and the other to the family Scolopendrellidae. The specimens were analysed with Genbank voucher specimens of Scutigerellidae as follows: Hanseniella sp. (Genbank accession # AF370839) and Scutigerellidae sp. FRL-2015 (Genbank accession #KP696391) as well as a Genbank voucher specimen of Scolopedrellidae, Symphylella sp. YG- 2006 (Genbank accession #EF576853). In addition, as local references, 26 reference specimens of Symphyla from the Pilbara were included in the analysis. Two specimens of centipede Scutigerina weberi (Genbank accession # DQ222166) and Scolopendra dawydoffi (Genbank accession #KR705635) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the two specimens from the Robe Valley, in addition to 29 reference specimens, placed the Robe Valley specimen in two distinct genetic lineages (Figure 8). The new Robe Valley lineages did not contain reference specimens and so were assigned to the new lineages SYM025 and SYM026 (Figure 8). Lineage SYM025 formed a poorly- supported clade with the Genbank specimen of Scutigerellidae and five specimens from Upper South Fortescue, Newman and Caramulla Creek in the Pilbara, three of which were assigned to the genus Scutigerella on the basis of morphology (Figure 8). Lineage SYM026 formed a well- supported clade with the Genbank specimen of Scolopendridae, and four specimens from Newman, Turner River, Yeelirrie Station and Boundary River in the Pilbara (Figure 8).

Differentiation within and between lineages Lineage SYM025 from Robe Valley differed from the reference specimens by between 15.0 and 28.1% sequence divergence and lineage SYM026 differed from the reference specimens by between 16.7 and 29.5% sequence divergence (Table 19).

Chilopoda Eleven specimens of Chilopoda from the Robe Valley (historical, Phase 3, Phase 1/2 repeat) were sequenced for COI (Table 20). Sequences were obtained from five specimens and these were analysed with eight Genbank vouchers from four families of Chilopoda: Cryptopidae, Geophilidae, Mecistocephalidae (in the superfamily Geophilomorpha) and Scolopendridae as follows: Cryptops japonicus (Genbank accession # AB610777), Cryptops pictus (Genbank accession #JF273290) Cormocephalus multidens (Genbank accession # KF676531), Scolopendra multidens (Genbank accession #AB614405), Gnathoribautia bonensis (Genbank accession # KF569297), Geophilus flavus (Genbank accession # JN306685), Stenotaenia linearis (Genbank accession # KR736251), and Mecistocephalus multidentatus (Genbank accession # AB610774) as well as 23 reference specimens of Chilopoda from 12 sites in the Pilbara. Two

2 Molecular systematics of subfauna from the Robe Valley

specimens of Onychophora, Metaperipatus inae (Genbank accession # HQ453464) and Opisthopatus cinctipes (Genbank accession #NC014273) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the five specimens from the Robe Valley in addition to 31 reference specimens, placed the Robe Valley specimens in five genetically distinct lineages (Figure 9). The specimens were not placed in lineages containing reference specimens, and were therefore assigned to the new lineages CHI021 – CHI025; Figure 9). Three of the lineages (CHI021 – CHI023) were placed in a well-supported clade with Genbank and Pilbara specimens of Cryptopidae (Figure 9). The remaining two lineages (CHI024, CHI025) were placed outside of the main clade containing the Pilbara and reference specimens of both Cryptopidae and Geophilomorpha, along with Genbank specimens of Scolopendridae (Figure 9).

Differentiation within and between lineages The five Chilopoda lineages from Robe Valley differed from one another by between 20.0 and 30.5% sequence divergence and from the reference specimens by between 16.3 and 29.8% sequence divergence (Table 21).

Coleoptera Reference specimens and outgroups Forty-two specimens of Coleoptera (historical, phase 3, phase 1/2 repeat) were sequenced for COI (Table 22). In order to reduce analysis time and to simplify the presentation of results, a preliminary neighbour-joining (NJ) analysis was conducted on the 42 Robe Valley Coleoptera specimens. Ten distinct genetic lineages were detected (Figure 10) and haplotypes were selected from each genetic lineage to include in a model-based phylogenetic analysis. Following a preliminary search on Genbank and the Helix database (consisting of 122 COI sequences of Coleoptera, including 23 provided by the WAM) for similar sequences, a reference data set was selected consisting of ten Genbank voucher sequences of Coleoptera as well as 40 reference specimens of Coleoptera from the Pilbara, including ten sequences provided by the WAM, selected for their similarity to the Robe Valley specimens. Two specimens of Hemiptera (Hemiptera sp. Genbank accession #GU671532) and Lycorma delicatula (Genbank accession #EU909203) were used as outgroups.

Phylogenetic analysis The phylogenetic analysis, which included the ten specimens from the Robe Valley in addition to 52 reference specimens, placed the Robe Valley specimens in ten genetically distinct lineages (Figure 11). Four of the ten lineages contained reference specimens and were assigned to the existing lineages CAN001 (Anthicidae), CCU004 (Curculionidae), CPT003 (Ptillidae), CST002 (Staphylinidae), whereas the six remaining lineages did not contain other reference specimens and were assigned to new lineages CCA012, CCA013 (Carabidae), CCU007 (Curculionidae), CUN 001, CUN002, CUN003 (unknown families; Figure 11). However, all of the new lineages did show significant relationships to reference specimens. Specifically, lineages CCA012 and CCA013 were placed in well-supported clades containing reference specimens of Carabidae and lineage CCU007 was placed in a well-supported clade containing reference specimens of Curculionidae, allowing tentative taxonomic assignments to those families (Figure 11). Lineages CUN001, CUN002 and CUN003 formed well-supported, but more distant relationships with GenBank specimens of Dermestidae, Anthicidae and Corylophidae, respectively, which may give starting points for future taxonomic investigation (Figure 11).

Differentiation within and between lineages

3 Molecular systematics of subfauna from the Robe Valley

The ten lineages of Coleoptera from the Robe Valley differed from one another by between 13.8 and 26.4% sequence divergence (Table 23). Distances between individuals within lineages averaged between 0 and 1.3% (Table 24). The Robe Valley lineages differed from the reference specimens by between 0 and 30.6% sequence divergence (Table 25). In particular, four Robe Valley lineages (IV234, IV405, IV392, IV422) differed from the nearest reference lineages by <1% (Table 25). IV232 differed from the nearest reference specimen by 3.7% and the remaining five lineages from Robe Valley differed from the reference specimens by > 10% (Table 25).

Conclusions COXI is widely considered to show suitable variation to distinguish species (Hebert et al., 2003a). In a comparison of COXI sequences for over 13,000 pairs of taxa, Hebert et al (2003b) found a mean of 11.1% sequence divergence between distinct species. Nearly 80% of the comparisons showed that species pairs differed from one another by greater than 8% sequence divergence. However, a taxon by taxon approach, examining the amount of phylogenetic variation within and between species is the most widely accepted method of defining species.

Symphyla Two distinct genetic lineages were detected at the Robe Valley, both of which differed from the reference specimens by >15%. This indicates that both lineages are likely to represent new species, which have so far not been detected in the Pilbara, based on the material available for comparison. One lineage, SYM025, appears to belong to the family Scutigerellidae, although the relationship was not well supported in the phylogenetic analysis. The second lineage, SYM026, is likely to belong to the Scolpendrellidae.

Chilopoda Five genetically distinct lineages of Chilopoda were detected at the Robe River Valley. The five lineages differed from one another by > 20% sequence divergence, thus each is likely to represent a distinct species. None of the five lineages showed close genetic relationships to reference specimens, differing from the references by >16.0% sequence divergence, thus the five species are likely to be new, based on the material available for comparison. Lineages CHI021 – CHI023 were placed with the Cryptopidae and appear to belong to that family. The remaining two lineages cannot be reliably assigned to a family, owing to poorly resolved relationships in the phylogenetic analysis, however, the sequence are most similar to specimens of Scolopendridae in the Helix and Genbank reference datasets.

Coleoptera Ten genetically distinct lineages of Coleoptera were detected at Robe Valley, differing from one another by >13%, indicating that each represents a distinct species. Four of the ten lineages (CCU004, CPT003, CST002 and CAN001) differed from the reference lineages by < 1%, indicating that they represent species that have been detected previously. It is worth noting that lineage CAN001 appears to be the introduced species Lasioderma serricorne, the cigarette beetle, which is a worldwide pest of tobacco and grain crops. The possibility that this species was a contaminant from field gear, vehicles, laboratories, etc, should be considered.

A fifth lineage (CCA012) differed from the nearest reference lineage (CCA001 from a previous survey in the Robe Valley) by 3.7%, suggesting that it is likely to belong to that previously- detected species, although further investigation may be required to resolve the relationship between the two lineages.

The remaining five lineages differed from the reference lineages by >10% sequence divergence, indicating that each is likely to represent a new species, which have so far not been detected in the Pilbara, based on the material available for comparison. However, we may have evidence to suggest taxonomic starting-points for these new lineages, based on significant relationships to Pilbara and Genbank reference specimens. In particular, lineages CCA013 and CCU007 appear to belong to the Carabidae and Curculionidae families, respectively, which are both common in the Pilbara, and lineage CUN002 appears to belong to

4 Molecular systematics of subfauna from the Robe Valley

the family Anthicidae, which has also been detected previously in the Pilbara, although we have limited specimens in our sequence database. In contrast, lineages CUN001 and CUN003 show more distant relationships to Genbank specimens of Dermestidae and Corylophidae, respectively. Both families are well-represented in Australia (Atlas of Living Australia).

Summary Seventeen lineages from three taxonomic groups were detected at Robe Valley, as shown below. The 17 lineages likely represent 17 species. Four or five of the species have been detected previously in the Pilbara, The remaining species at Robe Valley are likely to be new, based on the material available for comparison. One species of Coleoptera matched a Genbank sequence of an introduced species, which is a world-wide pest of grain and tobacco crops. The possibility that it was a contaminant from field gear, vehicles, laboratories, etc, should be considered.

Symphyla A single lineage was detected at Yandi, which differed from the reference specimens by >15%, indicating that it is a new species, based on the material available for comparison.

Scutigerellidae (putative): SYM025 = one species Scolopendridae: SYM026 = one species

Chilopoda Five lineages were detected at Robe Valley, likely corresponding to five species. None of the species have been detected previously in the Pilbara, based on the material available for comparison.

Cryptopidae: CHI021, CHI022, CHI023 = three species Scolopendridae (putative): CHI024, CHI025 = two species

Coleoptera Ten lineages were detected in the present study, corresponding to ten distinct species. Two species may belong to the Carabidae, two to the Curculionidae and one each to the Staphylinidae, Ptillidae, Anthicidae and Aniobidae. The remaining two species could not be placed into a family with confidence, however they showed significant relationships with families that are well represented in Australia, namely the Dermestidae and Corylophidae. Five species are likely to be new, based on the high genetic distances between them and the reference material available for comparison, whereas the remaining five appear to have been detected previously, although one species requires further investigation.

Carabidae: CCA012, CCA013 = two species Curculiondiae: CCU004, CCU007 = two species Anthicidae: CUN002 = one species Ptillidae: CPT003 = one species Staphylinidae: CST002 = one species Aniobidae: CAN001 = one species (introduced) Dermestide (putative): CUN001 = one species Corylophidae (putative): CUN3 = one species

References

Australian Faunal Directory. http://www.environment.gov.au/biodiversity/abrs/online- resources/. Accessed 27 Dec., 2015.

5 Molecular systematics of subfauna from the Robe Valley

Bayly, I. A.E, Ellis p. (1969). Haloniscus searlei chilton: An aquatic “terrestrial” isopod with remarkable powers of osmotic regulation. Comparative Biochemistry and Physiology 31: 523-528

Drummond AJ, Ashton B, Buxton S, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M, Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A (2011) Geneious v5.4, http://www.geneious.com/ Harvey, M. S., Berry, O. Edward, K. L., Humphreys, G. (2008) Molecular and morphological systematics of hypogean schizomids (Schizomida: Hubbardiidae) in semiarid Australia. Invertebrate Systematics 22: 167-194.

6 Molecular systematics of subfauna from the Robe Valley

Table 18. Specimens of Symphyla used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat.

Specimen ID Drillhole ID Provenance Helix ID Lineage SSp-2010-130 COBRC0030 H IV348 Failed repeat PCR SSp-2010-131 COBRC0003 H IV349 Failed repeat PCR SSp-2010-198 COBRC0009 H IV350 NoAmp SSp-2010-218 COBRC0010 H IV351 contam SSp-2010-229 COBRC0033 H IV352 Failed repeat PCR SSp-2010-69 COBRC0015 H IV353 Failed repeat PCR SSp-2010-84 TOBRC0038 H IV354 SYM025 GR15MEB0003-20160317-T3-03 GR15MEB0003 P3 IV271 NoAmp 138524 MEARC4273 P 1/2 IV355 SYM026

Table 19. (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Symphyla detected at Robe Valley and the reference lineages as shown in Figure 8.

Table 20. Specimens of Chilopoda used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat.

Specimen ID Drillhole ID Provenance Helix ID Lineage SSp-2010-325 COBRC0019 H IV347 Failed repeat PCR 93283 TBRC038 H 93283 NoAmp 93287 TBRC036 H 93287 NoAmp 93778 TBRC141 H 93778 NoAmp 93780 KBRC096 H 93780 NoAmp 98714 UCRC055 H 98714 NoAmp 120191 H 120191 CHI021 132678 H 132678 CHI022 93779 TBRC029 H 93779 CHI024 RC14MEB0115-20160317-T2-02 RC14MEB0115 P3 IV231 CHI023 138572 DD14MEL0001 P1/2 re IV142 CHI025

Table 21. (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Chilopoda detected at Robe Valley and the reference lineages as shown in Figure 9.

7 Molecular systematics of subfauna from the Robe Valley

Table 22. Specimens of Coleoptera used in the present study and the lineage to which they were assigned, based on variation at the COXI gene. nd=no data. Provenance is classified as either historical (H), by phase (P) or as provided by the WAM (WAM); re=repeat.

Specimen ID Drillhole ID Provenance Cap No. Helix ID Lineage MEK1478P7T2-2 MEK1478 H 44 IV395 CCU007 MEK1478P7T2-2 MEK1478 H 44 IV396 CCU007 MEK1718P7T3-2 MEK1718 H 216 IV397 CCU007 MEK1718P7T3-2 MEK1718 H 216 IV398 CCU007 MEK1718P7T3-2 MEK1718 H 216 IV399 CCU007 MEK1721P7T1-3 MEK1721 H 147 IV400 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV401 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV402 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV403 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV404 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV405 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV406 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV407 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV408 CPT003 MEK1721P7T1-3 MEK1721 H 147 IV409 CPT003 K0968P7T3-3 K0968 H 100 IV410 CPT003 MEGRC0495P6T2-3 MEGRC0495 H 166 IV411 CUN003 MEGRC0495P6T2-3 MEGRC0495 H 166 IV412 CUN003 MEGRC0097P6T1-5 MEGRC0097 H 112/172? IV413 CUN003 MEARC2657P1T1-3 MEARC2657 H 133 IV414 CCU004 RC08SILV0683p3t1-1 RC08SILV0683 H 286 IV415 CCA013 D08BU075P1T1-4 D08BU075 H 14 black IV416 CUN001 D08BU075P1T1-4 D08BU075 H 14 black IV417 CUN001 D08BU075P1T1-4 D08BU075 H 14 black IV418 CUN001 D08BU075P1T1-4 D08BU075 H 14 black IV419 CUN001 D08BU075P1T1-4 D08BU075 H 14 black IV420 CUN001 D08BU075P1T1-4 D08BU075 H 14 black IV421 CUN001 E004FP3T1-4 E004F H 14 red IV422 CAN001 BUNW01114P1T2-3 BUNW01114 H 6 IV423 CUN002 RHRC013P1T3-3 RHRC013 H 19 IV424 CAN001 RC15MEB0107-20160317-T2-02 RC15MEB0107 P3 133 IV232 CCA012 RC15MEC0192-20160316-T2-03 RC15MEC0192 P3 197 IV233 CCU004 RC15MEC0192-20160316-T3-03 RC15MEC0192 P3 200 IV234 CCU004 RC15MEC0192-20160316-T3-04 RC15MEC0192 P3 201 IV235 CCU004 MEARC5093-20151001-T1-03 MEARC5093 P1/2 re 48 IV388 CUN003 MEARC5093-20151001-T2-02 MEARC5093 P1/2 re 51 IV389 CST002 DD14MEC0006T2-02 DD14MEC0006 P1/2 re 791 IV390 CST002 DD14MEC0006T2-02 DD14MEC0006 P1/2 re 791 IV391 CST002 DD14MEC0006T2-02 DD14MEC0006 P1/2 re 791 IV392 CST002 DD14MEC0006T2-02 DD14MEC0006 P1/2 re 791 IV393 CST002 DD14MEC0006T2-02 DD14MEC0006 P1/2 re 791 IV394 CST002 IV149 RC14MEB0068 P1/2 re Failed PCR IV149 CCU004

8 Molecular systematics of subfauna from the Robe Valley

Table 23. Mean genetic distance (below diagonal) and standard error (above diagonal, in blue) between lineages of Coleoptera detected in the NJ analysis as shown in Figure 10.

Lineage 1 2 3 4 5 6 7 8 9 10 1 0.015 0.016 0.015 0.017 0.017 0.016 0.016 0.018 0.018 2 0.204 0.014 0.015 0.015 0.015 0.016 0.015 0.018 0.017 3 0.207 0.138 0.017 0.016 0.016 0.016 0.016 0.016 0.016 4 0.208 0.178 0.171 0.017 0.015 0.015 0.016 0.018 0.017 5 0.209 0.195 0.198 0.204 0.014 0.015 0.015 0.017 0.018 6 0.218 0.186 0.190 0.193 0.186 0.014 0.013 0.015 0.017 7 0.222 0.201 0.203 0.186 0.208 0.186 0.016 0.017 0.017 8 0.227 0.227 0.225 0.223 0.212 0.205 0.213 0.014 0.014 9 0.263 0.233 0.239 0.264 0.250 0.238 0.263 0.245 0.011 10 0.261 0.245 0.237 0.261 0.257 0.242 0.259 0.256 0.131

Table 24. Mean distance (D) and standard error (s.e.) within lineages of Coleoptera detected in the NJ analysis as shown in Figure 10 and the number (N) of individuals assigned to that lineage. n/c = not calculated, for groups where n=1. Rep=individual selected to represent the lineage in the model-based analysis.

Lineage D s.e. N rep 1 0.001 0.001 10 IV405 2 n/c n/c 1 IV232 3 n/c n/c 1 IV415 4 0.001 0.001 2 IV422 5 0.000 0.000 6 IV419 6 n/c n/c 1 IV423 7 0.003 0.001 6 IV392 8 0.006 0.002 4 IV413 9 0.013 0.003 4 IV234 10 0.001 0.001 5 IV395

Table 25. (attached). Genetic distances (below diagonal) and standard error (above diagonal, in blue) between specimens of Coleoptera detected at Robe Valley and the reference lineages as shown in Figure 11.

9 Molecular systematics of subfauna from the Robe Valley

Figure 8. Bayesian analysis of COXI haplotypes of Symphyla from the present study and reference specimens from previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site. DQ222166 Scutigerina weberi Scutigeridae KR705635 Scolopendra dawydoffi Scolopendridae IV354 SSp 2010 84 Symphyella sp. B9 SYM025 0.61 G121 Symphyla Upper S. Fortescue SYM015 KP696391 Scutigerellidae sp. FRL 2015

1.00 EO20 Scutigerella B09 EXR1462R Newman SYM006 1.00 EXR1356 Caramulla Creek SYM006 0.61 1.00 EO28 Scutigerella sp. B06 EXR0641 Newman SYM020 1.00 EO31 Scutigerella sp. B06 EXR0641 Newman SYM020 GH23 Hanseniella Han PES16403 SYM023 0.97 JIN0290 Hancock Range South SYM005 0.60 EJUNK02 Wheelarra Hill SYM007 1.00 1.00 PI059 Wheelarra Hill SYM003 0.53 LB069 Wheelarra Hill SYM004 0.98 G368 Symphyla sp MA LN8697 SYM017

0.93 G366 Symphyla sp OP LN9204 SYM016 0.99 J7 22 Wheelarra Hill SYM008 0.91 JD04 DD11YOX32020151124 T2 3 SYM001 EO23 Hanseniella sp. B19 EES0155 SYM021 1.00 1.00 EO24 Hanseniella sp. B19 EA0110RT SYM021 0.61 IS72 KBRC1475 20150708 T2 02 Symphyla SYM024 EO26 Hanseniella sp. B20 Now sp. EB0266R SYM022 AF370839 Hanseniella sp. Scutigerillidae 1.00 G42 Symphyla SYM010 1.00 G44 Symphyla SYM011 1.00 1.00 1.00 G45 Symphyla SYM012 G46 Symphyla SYM013 IV355 138524 Symphyla SYM026 0.61 G43 Symphyla Yeelirrie Station SYM014

1.00 MUDUNK0 J14 4 S Boundary River sp B4 SYM009 EF576853 Symphylella sp. YG 2006 Scolopendrellidae 1.00 1.00 G494 111138b Symphyla Turner River SYM018 EXR0425 Newman SYM002 0.1

10 Molecular systematics of subfauna from the Robe Valley

Figure 9. Neighbour-joining analysis of specimens of Chilopoda from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

EF624055 Metaperipatus inae NC014273 Opisthopatus cinctipes 120191 Chilopoda Cryptopidae CHI021 0.67 0.99 G124 Cryptops Upper South Fortescue CHI004 IV231 RC14MEB0115 20160317 T2 02 CHI023 0.54 BQ12 PES 5087 Cryptops won1 CHI007 0.94 0.51 G349 Cryptopidae sp MJ LN9808 CHI005 DF14 9703 Cryptopidae sp. CHI016 0.91 FH15 nr Cryptops sp B34 DL0414 CHI018 CQ5 nr Cryptops sp. B11 PCRC088 CHI012 0.830.88 FH16 nr Cryptops sp B34 EW0769 CHI019 AB610777 Cryptops japonicus 0.831.00 IV143 MEARC4383 20151001 T2 01 CHI002 CQ2 nr Cryptops sp. B15 CCWUNK05 CHI013 Cryptopidae 132678 Chilopoda Cryptopidae CHI022 1.00 JF273290 Cryptops pictus 1.00 BX11 PES 0968 Cryptops pilbara1 CHI011 1.00 1.00 EY5 Cryptops sp B35 CHI017 BX12 PES 3783 Cryptops pilbara2 CHI009 0.99 0.98 CQ6 Cryptops sp. B30 1106 13 EX13 CHI014 0.54 BX13 991 5262 Cryptops MH1 CHI010 1.00 1.00 BX8 PES 5889 Cryptops MH2 CHI010 0.59 1.00 CQ3 Cryptops sp. B32 CC1798 CHI015 1.00 BX9 PES 5272 Cryptops MH2 CHI008 AK10 PE111022 Cryptopidae DC10 CHI006 AB610774 Mecistocephalus multidentatus 1.00 IS87 KBRC136220150911T2 03 Geophilida sp CHI020 1.00 IV139 DD14MRR0004T1 01 CHI001 0.99 1.00 1.00 IV140 DD14MRR0004T1 01 CHI001 Geophilomorpha 1.00 IV141 DD14MRR0005 T1 04 CHI001 JN306685 Geophilus flavus 1.00 0.99 1.00 KR736251 Stenotaenia linearis KF569297 Gnathoribautia bonensis 0.99 AB614405 Scolopendra multidens IV142 138572 Chilopoda CHI025 0.63 G123 Cryptops Upper South Fortescue CHI003 Scolopendridae? 1.00 KF676531 Cormocephalus hartmeyeri Scolopendridae 93779 Chilopoda Cryptopidae sp. CHI024 0.1

11 Molecular systematics of subfauna from the Robe Valley

Figure 10. Neighbour-joining analysis of specimens of Coleoptera from the present study. Numbers on major nodes correspond to bootstrap support over 100 iterations; values <50% are not shown. Scale bar= genetic distance. The specimens used to represent each lineage in the model-based phylogenetic analysis are highlighted in yellow.

IV408 MEK1721P7T1-3 Coleoptera IV410 K0968P7T3-3 Coleoptera IV407 MEK1721P7T1-3 Coleoptera IV406 MEK1721P7T1-3 Coleoptera

100 IV405 MEK1721P7T1-3 Coleoptera IV404 MEK1721P7T1-3 Coleoptera Gp 1 IV403 MEK1721P7T1-3 Coleoptera 100 IV402 MEK1721P7T1-3 Coleoptera IV401 MEK1721P7T1-3 Coleoptera IV400 MEK1721P7T1-3 Coleoptera 26 IV409 MEK1721P7T1-3 Coleoptera

93 IV232 RC15MEB0107-20160317-T2-02 Coleoptera Gp 2 IV415 RC08SILV0683p3t1-1 Coleoptera Gp 3 15 52 IV422 E004FP3T1-4 Coleoptera Gp 4 100 IV424 RHRC013P1T3-3 Coleoptera IV416 D08BU075P1T1-4 Coleoptera IV417 D08BU075P1T1-4 Coleoptera 100 40 IV419 D08BU075P1T1-4 Coleoptera 30 Gp 5 IV420 D08BU075P1T1-4 Coleoptera 18 23 IV418 D08BU075P1T1-4 Coleoptera 100 IV421 D08BU075P1T1-4 Coleoptera IV423 BUNW01114P1T2-3 Coleoptera Gp 6 IV390 DD14MEC0006T2-02 Coleoptera 32 IV389 MEARC5093-20151001-T2-02 Coleoptera 100 68 IV391 DD14MEC0006T2-02 Coleoptera Gp 7 71 IV392 DD14MEC0006T2-02 Coleoptera 53 IV393 DD14MEC0006T2-02 Coleoptera 59 IV394 DD14MEC0006T2-02 Coleoptera IV388 MEARC5093-20151001-T1-03 Coleoptera IV413 MEGRC0097P6T1-5 Coleoptera 100 Gp 8 99 IV411 MEGRC0495P6T2-3 Coleoptera 77 IV412 MEGRC0495P6T2-3 Coleoptera

61 IV233 RC15MEC0192-20160316-T2-03 Coleoptera 57 IV234 RC15MEC0192-20160316-T3-03 Coleoptera 100 IV149 RC14MEB0068 Coleoptera Gp 9 100 IV235 RC15MEC0192-20160316-T3-04 Coleoptera IV414 MEARC2657P1T1-3 Coleoptera 100 IV397 MEK1718P7T3-2 Coleoptera IV398 MEK1718P7T3-2 Coleoptera 100 IV395 MEK1478P7T2-2 Coleoptera Gp 10 61 IV396 MEK1478P7T2-2 Coleoptera 57 IV399 MEK1718P7T3-2 Coleoptera

0.02

12 Molecular systematics of subfauna from the Robe Valley

Figure 11. Bayesian analysis of COXI haplotypes of Coleoptera from the present study and reference specimens from Genbank and previous surveys in the Pilbara. Numbers on major nodes correspond to posterior probabilities; values <50% are not shown. Specimens from the present study are highlighted in yellow; GenBank voucher specimens highlighted in turquoise. Scale bar= number of substitutions per site. 001 EU909203 Lycorma delicatula GU671532 Hemiptera sp. AX43 ID29P1T1 3 CST001 WAME89084 Staphylinidae sp. 1 1.00 1.00 1.00 WAME89094 Staphylinidae sp. 1 1.00 WAME89085 Staphylinidae sp. 1 WAME89101 Staphylinidae sp. 1 1.00 0.96 WAME89111 Staphylinidae sp. 1 Staphylinidae IE25 CBRC122P3T2 3 CST002 1.00 1.00 0.92 IV392 DD14MEC0006T2 02 CST002 1.00 KP422158 Pselaphinae sp. NZAC Staphylinidae JV04 RC15BS40051 20160608 T3 01B CST003 JV05 RC15BS4B0415 20160609 T2 01 CST004 BX6 PES 5290 Bembidion MH1 CCA008 1.00 BX7 PES 5285 Bembidion MH1 CCA008 G202 Anillini 100490 CCA004 0.85 1.00 0.84 G436 110879 Anillini sp CCA004 0.74 1.00 G122 Anillini CCA003 G203 Anillini 107877 CCA005 IE27 CWRC179P4T3 5 CCA010 1.00 Carabidae 1.00 IE28 CWRC274P4T3 5 CCA011 CA4 11 1461 Anillini sp. McP1 CCA009 G216 Coleoptera 109656 CCA006 0.98 IV151 MEARC3814 20150807 T1 02 CCA001 0.75 1.00 0.97 0.52 IV232 RC15MEB0107 20160317 T2 02 CCA012 LN6119 CA0021 CCA002 0.95 G437 110722 Anillini sp CCA007 IV415 RC08SILV0683p3t1 1 CCA013 1.00 KM447718 Harpalus tardus Carabidae AX62 11DDH051P1T2 3 CUN001 IE15 BHRC122 20141216 T2 01 CPT002 1.00 IE17 BHRC058 20141215 T1 01 CPT003 0.98 Ptillidae 0.96 IV405 MEK1721P7T1 3 CPT003 1.00 1.00 KU519734 WAME89168 Ptiliidae sp. 1 IV147 DD14MRR0004T1 03 CPT002 0.65 IV422 E004FP3T1 4 CAN001 1.00 Aniobidae 0.98 KJ680546 Lasioderma serricorne Aniobidae 0.68 JN171062 Bembidion lugubre Carabidae JV03 RC15BS40051 20160608 T3 01A Coleoptera IV419 D08BU075P1T1 4 CUN001 1.00 KM578824 Dermestes frischii Dermestidae Dermestidae 0.93 FH1 Curculionidae Genus2 B15 EW0540 CCU003 1.00 1.00 WAME89155 G114 Curculionidae CCU001 GU213687 Echinodera ibleiensis Curculionidae IV150 RC14MEB0060 20151001 T2 02 CCU004 0.66 1.00 0.99 IV234 RC15MEC0192 20160316 T3 03 CCU004 1.00 1.00 IV152 MEARC4400 t1 01 CCU005 Curculionidae 1.00 1.00 1.00 WAME89188 Curculionidae sp. 8 0.91 1.00 KU519717 WAME89186 Curculionidae sp. 8 IV395 MEK1478P7T2 2 CCU007 HE615891 Trigonopterus sp. Curculionidae 0.99 WAME89176 Curculionidae sp. 5 0.99 G115 Curculionidae CCU002

1.00 IV153 MEARC4400 t1 01 CAN003 0.54 KJ962132 Stricticollis tobias Anthicidae Anthicidae 1.00 KM439906 Anthicus flavipes Anthicidae 1.00 IV423 BUNW01114P1T2 3 CUN002 IE19 BHRC218 20141216 T2 01

1.00 IE30 MEBRC0021 0712 T1 2 Latridiidae? 1.00 JV01 GR14BS40032 20160608 T1 01 Latridiidae? 0.97 JV02 RC11BS3045 20160609 T1 01 Coleoptera 1.00 IV413 MEGRC0097P6T1 5 CUN003 KM439290 Orthoperus nigrescens Corylophidae Corylophidae

0.1

13 Table 19. Estimates of Evolutionary Divergence between Sequences

Lineage IV354 IV355 Han-sp. Sym-sp. Scu-sp. EJUNK02 EO20 EO23 EO24 EO26 EO28 EO31 EXR0425 EXR1356 G121 G366 G368 G42 G43 G44 G45 G46 G494 GH23 IS72 IV354 SSp-2010-84 Symphyella sp. B9 0.016 0.013 0.014 0.013 0.014 0.013 0.013 0.014 0.015 0.013 0.013 0.015 0.014 0.014 0.014 0.015 0.017 0.017 0.016 0.013 0.020 0.015 0.015 0.012 IV355 138524 Symphyla 0.224 0.018 0.016 0.017 0.015 0.016 0.017 0.017 0.015 0.017 0.017 0.015 0.017 0.017 0.016 0.017 0.019 0.014 0.018 0.018 0.021 0.015 0.017 0.016 AF370839 Hanseniella Scutigerillidae 0.221 0.262 0.015 0.014 0.015 0.013 0.015 0.015 0.016 0.013 0.013 0.016 0.012 0.016 0.013 0.015 0.015 0.018 0.016 0.014 0.020 0.015 0.015 0.013 EF576853 Symphylella sp. YG-2006 0.250 0.194 0.281 0.014 0.015 0.014 0.016 0.016 0.017 0.015 0.015 0.015 0.014 0.017 0.015 0.016 0.017 0.015 0.018 0.015 0.019 0.014 0.016 0.014 KP696391 Scutigerellidae sp. FRL-2015 0.204 0.270 0.220 0.264 0.014 0.013 0.016 0.016 0.016 0.014 0.014 0.016 0.013 0.015 0.015 0.015 0.017 0.019 0.019 0.015 0.020 0.016 0.014 0.013 EJUNK02 BHP1154 Jimblebar E 0.186 0.256 0.209 0.284 0.215 0.015 0.016 0.015 0.015 0.015 0.015 0.017 0.015 0.014 0.016 0.015 0.019 0.020 0.016 0.015 0.021 0.017 0.015 0.015 EO20 Hanseniella sp. B04 NOW scutigerella B09 EXR1462R 0.163 0.247 0.226 0.253 0.195 0.192 0.013 0.013 0.015 0.013 0.013 0.016 0.003 0.014 0.012 0.014 0.017 0.019 0.016 0.014 0.019 0.014 0.013 0.014 EO23 Hanseniella sp. B19 EES0155 0.176 0.217 0.223 0.252 0.211 0.197 0.181 0.006 0.015 0.013 0.013 0.016 0.013 0.014 0.014 0.015 0.016 0.017 0.019 0.015 0.022 0.014 0.015 0.014 EO24 Hanseniella sp. B19 EA0110RT 0.185 0.216 0.218 0.249 0.216 0.194 0.183 0.027 0.015 0.014 0.014 0.016 0.013 0.015 0.014 0.016 0.016 0.016 0.019 0.016 0.022 0.014 0.015 0.015 EO26 Hanseniella sp. B20 Now sp. EB0266R 0.194 0.225 0.240 0.263 0.229 0.228 0.181 0.173 0.168 0.015 0.015 0.015 0.015 0.015 0.015 0.015 0.016 0.017 0.018 0.015 0.020 0.015 0.014 0.014 EO28 Scutigerella sp. B06 EXR0641 0.177 0.243 0.217 0.262 0.207 0.195 0.125 0.176 0.180 0.197 0.000 0.017 0.014 0.015 0.013 0.014 0.016 0.018 0.017 0.015 0.020 0.014 0.014 0.015 EO31 Scutigerella sp. B06 EXR0641 0.177 0.243 0.217 0.262 0.207 0.195 0.125 0.176 0.180 0.197 0.000 0.017 0.014 0.015 0.013 0.014 0.016 0.018 0.017 0.015 0.020 0.014 0.014 0.015 EXR0425 BHP1183 0.249 0.224 0.269 0.217 0.270 0.273 0.261 0.250 0.249 0.243 0.270 0.270 0.016 0.014 0.016 0.016 0.017 0.017 0.019 0.016 0.021 0.014 0.017 0.014 EXR1356 BHP1222 Caramulla Creek 0.169 0.245 0.226 0.250 0.195 0.191 0.008 0.183 0.183 0.180 0.133 0.133 0.258 0.014 0.013 0.014 0.018 0.019 0.016 0.014 0.020 0.014 0.014 0.014 G121 Symphyla 0.159 0.233 0.216 0.246 0.216 0.205 0.177 0.183 0.180 0.186 0.177 0.177 0.252 0.176 0.012 0.013 0.018 0.016 0.017 0.014 0.021 0.014 0.015 0.015 G366 Symphyla sp OP LN9204 0.174 0.217 0.221 0.252 0.211 0.190 0.178 0.162 0.164 0.177 0.171 0.171 0.244 0.180 0.174 0.013 0.017 0.018 0.019 0.015 0.020 0.014 0.015 0.014 G368 Symphyla sp MA LN8697 0.195 0.228 0.220 0.258 0.211 0.202 0.209 0.184 0.188 0.190 0.196 0.196 0.266 0.212 0.185 0.175 0.017 0.017 0.017 0.015 0.018 0.017 0.016 0.015 G42 Symphyla 0.239 0.261 0.216 0.230 0.221 0.260 0.224 0.219 0.212 0.215 0.228 0.228 0.252 0.220 0.241 0.226 0.239 0.015 0.016 0.017 0.017 0.016 0.016 0.014 G43 Symphyla 0.244 0.181 0.252 0.211 0.261 0.256 0.251 0.233 0.225 0.231 0.251 0.251 0.217 0.244 0.235 0.227 0.225 0.240 0.018 0.017 0.021 0.014 0.019 0.018 G44 Symphyla 0.209 0.264 0.208 0.252 0.247 0.218 0.226 0.228 0.221 0.221 0.245 0.245 0.263 0.224 0.207 0.202 0.224 0.187 0.246 0.013 0.015 0.017 0.016 0.016 G45 Symphyla 0.218 0.253 0.200 0.267 0.238 0.236 0.222 0.213 0.206 0.219 0.231 0.231 0.288 0.224 0.215 0.206 0.220 0.182 0.268 0.118 0.015 0.016 0.014 0.015 G46 Symphyla 0.251 0.295 0.217 0.266 0.251 0.240 0.232 0.244 0.248 0.235 0.216 0.216 0.294 0.235 0.242 0.252 0.234 0.215 0.273 0.178 0.158 0.019 0.021 0.021 G494 111138b Symphyla 0.281 0.223 0.273 0.194 0.282 0.282 0.271 0.256 0.253 0.256 0.271 0.271 0.258 0.269 0.265 0.260 0.280 0.240 0.200 0.257 0.275 0.269 0.015 0.014 GH23 Hanseniella Han-PES16403 0.186 0.255 0.219 0.261 0.217 0.189 0.170 0.180 0.179 0.181 0.182 0.182 0.263 0.176 0.178 0.185 0.190 0.224 0.252 0.219 0.227 0.252 0.278 0.012 IS72 KBRC1475 20150708 T2 02 Symphyla 0.150 0.234 0.221 0.252 0.208 0.205 0.170 0.168 0.163 0.179 0.187 0.187 0.250 0.177 0.197 0.196 0.217 0.207 0.257 0.229 0.210 0.241 0.277 0.181 J7-22 Wheelarra Hill 0.173 0.215 0.241 0.258 0.198 0.202 0.191 0.162 0.165 0.170 0.185 0.185 0.241 0.191 0.171 0.169 0.185 0.254 0.238 0.232 0.223 0.274 0.287 0.175 0.197 JD04 DD11YOX32020151124-T2-3 Symphyla 0.185 0.235 0.214 0.272 0.231 0.207 0.184 0.201 0.187 0.195 0.189 0.189 0.283 0.182 0.182 0.183 0.195 0.241 0.252 0.217 0.218 0.265 0.293 0.202 0.199 JIN0290 BHP779 Hancock Range South 0.208 0.253 0.229 0.276 0.217 0.211 0.200 0.192 0.195 0.226 0.203 0.203 0.276 0.197 0.199 0.186 0.202 0.212 0.262 0.241 0.224 0.269 0.285 0.196 0.197 LB069 BHP984 Hashimoto 0.196 0.265 0.199 0.288 0.219 0.029 0.193 0.193 0.192 0.235 0.201 0.201 0.270 0.192 0.209 0.184 0.201 0.258 0.260 0.228 0.240 0.244 0.277 0.194 0.199 MUDUNK0 J14-4 S Boundary R sp B4 0.264 0.167 0.255 0.191 0.263 0.246 0.255 0.231 0.224 0.260 0.247 0.247 0.220 0.252 0.242 0.243 0.269 0.260 0.194 0.274 0.277 0.284 0.217 0.272 0.263 PI059 BHP935 Hashimoto 0.185 0.256 0.209 0.284 0.214 0.002 0.191 0.195 0.195 0.228 0.194 0.194 0.273 0.189 0.203 0.189 0.200 0.260 0.256 0.218 0.236 0.240 0.282 0.189 0.203 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. employers beliableforanydamages,includingbutnotlimitedtospecial, consequential,orotherdamages. textforaspecificpurpose,use,orapplication. Authors specificallydisclaimallotherwarrantiesexpressedorimplied, includingbutnotlimitedtothedeterminationofsuitabilitythiscaption Disclaimer: Although utmostcarehasbeentakentoensurethecorrectnessofcaption,captiontextisprovided"asis"withoutanywarrantykind. anyerrorsorproblemstotheauthorsimmediately(www.megasoftware.net).Authors advisetheusertocarefullycheckcaptionprioritsuseforanypurposeandreport Innoeventshallthe authorsandtheir The numberofbasedifferences persitefrombetweensequencesareshown.Standarderrorestimate(s)shownabovethediagonal. The analysisinvolved42nucleotide sequences.Codonpositionsincludedwere1st+2nd+3rd+Noncoding. were removedforeach All ambiguouspositions sequence pair. There wereatotalof495positionsinthefinaldataset.Evolutionaryanalyses wereconductedinMEGA6[1]. .1 .1 .1 .1 .1 0.014 0.014 0.015 0.014 0.015 0.013 .1 .1 .1 .1 .1 0.015 0.016 0.016 0.018 0.017 0.015 .1 .1 .1 .1 .1 0.015 0.016 0.015 0.013 0.016 0.015 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.015 0.017 0.015 .1 .1 .1 .1 .1 0.014 0.015 0.014 0.014 0.015 0.014 .1 .1 .1 .0 .1 0.002 0.015 0.006 0.015 0.015 0.015 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.015 0.016 0.014 .1 .1 .1 .1 .1 0.016 0.016 0.015 0.013 0.017 0.014 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.013 0.017 0.015 .1 .1 .1 .1 .1 0.015 0.016 0.016 0.015 0.016 0.015 .1 .1 .1 .1 .1 0.014 0.015 0.014 0.014 0.015 0.014 .1 .1 .1 .1 .1 0.014 0.015 0.014 0.014 0.015 0.014 .1 .1 .1 .1 .1 0.017 0.014 0.017 0.015 0.018 0.015 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.015 0.016 0.013 .1 .1 .1 .1 .1 0.014 0.016 0.014 0.015 0.015 0.013 .1 .1 .1 .1 .1 0.016 0.015 0.016 0.014 0.014 0.015 .1 .1 .1 .1 .1 0.015 0.017 0.015 0.015 0.014 0.015 .1 .1 .1 .1 .1 0.019 0.017 0.018 0.016 0.018 0.018 .1 .1 .1 .2 .1 0.020 0.015 0.020 0.017 0.019 0.017 .1 .1 .1 .1 .1 0.016 0.018 0.016 0.016 0.018 0.018 .1 .1 .1 .1 .1 0.015 0.015 0.014 0.015 0.016 0.014 .2 .2 .2 .2 .1 0.021 0.019 0.021 0.022 0.023 0.022 .1 .1 .1 .1 .1 0.017 0.016 0.016 0.014 0.016 0.015 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.015 0.015 0.014 .1 .1 .1 .1 .1 0.015 0.015 0.015 0.014 0.017 0.015 0.178 .0 0.222 0.205 .1 .1 0.195 0.218 0.212 .5 .5 .7 0.244 0.275 0.254 0.250 .0 .0 .0 .2 0.244 0.028 0.209 0.207 0.200

J7-22 .1 .1 .1 .1 0.014 0.015 0.016 0.015 0.015

JD04 .1 .1 .1 0.015 0.015 0.015 0.016

JIN0290 .1 .1 0.014 0.015 0.014

LB069 .1 0.006 0.015

J14-4 0.015

PI059 Table 19. Estimates of Evolutionary Divergence between Sequences

Lineage IV354 IV355 Han-sp. Sym-sp. Scu-sp. EJUNK02 EO20 EO23 EO24 EO26 EO28 EO31 EXR0425 EXR1356 G121 G366 G368 G42 G43 G44 G45 G46 G494 GH23 IS72 J7-22 JD04 JIN0290 LB069 J14-4 PI059 IV354 SSp-2010-84 Symphyella sp. B9 0.016 0.013 0.014 0.013 0.014 0.013 0.013 0.014 0.015 0.013 0.013 0.015 0.014 0.014 0.014 0.015 0.017 0.017 0.016 0.013 0.020 0.015 0.015 0.012 0.013 0.015 0.014 0.015 0.014 0.014 IV355 138524 Symphyla 0.224 0.018 0.016 0.017 0.015 0.016 0.017 0.017 0.015 0.017 0.017 0.015 0.017 0.017 0.016 0.017 0.019 0.014 0.018 0.018 0.021 0.015 0.017 0.016 0.015 0.017 0.018 0.016 0.016 0.015 AF370839 Hanseniella Scutigerillidae 0.221 0.262 0.015 0.014 0.015 0.013 0.015 0.015 0.016 0.013 0.013 0.016 0.012 0.016 0.013 0.015 0.015 0.018 0.016 0.014 0.020 0.015 0.015 0.013 0.015 0.016 0.013 0.015 0.016 0.015 EF576853 Symphylella sp. YG-2006 0.250 0.194 0.281 0.014 0.015 0.014 0.016 0.016 0.017 0.015 0.015 0.015 0.014 0.017 0.015 0.016 0.017 0.015 0.018 0.015 0.019 0.014 0.016 0.014 0.015 0.017 0.015 0.015 0.015 0.015 KP696391 Scutigerellidae sp. FRL-2015 0.204 0.270 0.220 0.264 0.014 0.013 0.016 0.016 0.016 0.014 0.014 0.016 0.013 0.015 0.015 0.015 0.017 0.019 0.019 0.015 0.020 0.016 0.014 0.013 0.014 0.015 0.014 0.014 0.015 0.014 EJUNK02 BHP1154 Jimblebar E 0.186 0.256 0.209 0.284 0.215 0.015 0.016 0.015 0.015 0.015 0.015 0.017 0.015 0.014 0.016 0.015 0.019 0.020 0.016 0.015 0.021 0.017 0.015 0.015 0.015 0.015 0.015 0.006 0.015 0.002 EO20 Hanseniella sp. B04 NOW scutigerella B09 EXR1462R 0.163 0.247 0.226 0.253 0.195 0.192 0.013 0.013 0.015 0.013 0.013 0.016 0.003 0.014 0.012 0.014 0.017 0.019 0.016 0.014 0.019 0.014 0.013 0.014 0.014 0.016 0.015 0.015 0.015 0.015 EO23 Hanseniella sp. B19 EES0155 0.176 0.217 0.223 0.252 0.211 0.197 0.181 0.006 0.015 0.013 0.013 0.016 0.013 0.014 0.014 0.015 0.016 0.017 0.019 0.015 0.022 0.014 0.015 0.014 0.014 0.017 0.013 0.015 0.016 0.016 EO24 Hanseniella sp. B19 EA0110RT 0.185 0.216 0.218 0.249 0.216 0.194 0.183 0.027 0.015 0.014 0.014 0.016 0.013 0.015 0.014 0.016 0.016 0.016 0.019 0.016 0.022 0.014 0.015 0.015 0.015 0.017 0.013 0.015 0.015 0.015 EO26 Hanseniella sp. B20 Now sp. EB0266R 0.194 0.225 0.240 0.263 0.229 0.228 0.181 0.173 0.168 0.015 0.015 0.015 0.015 0.015 0.015 0.015 0.016 0.017 0.018 0.015 0.020 0.015 0.014 0.014 0.015 0.016 0.015 0.016 0.016 0.015 EO28 Scutigerella sp. B06 EXR0641 0.177 0.243 0.217 0.262 0.207 0.195 0.125 0.176 0.180 0.197 0.000 0.017 0.014 0.015 0.013 0.014 0.016 0.018 0.017 0.015 0.020 0.014 0.014 0.015 0.014 0.015 0.014 0.014 0.015 0.014 EO31 Scutigerella sp. B06 EXR0641 0.177 0.243 0.217 0.262 0.207 0.195 0.125 0.176 0.180 0.197 0.000 0.017 0.014 0.015 0.013 0.014 0.016 0.018 0.017 0.015 0.020 0.014 0.014 0.015 0.014 0.015 0.014 0.014 0.015 0.014 EXR0425 BHP1183 0.249 0.224 0.269 0.217 0.270 0.273 0.261 0.250 0.249 0.243 0.270 0.270 0.016 0.014 0.016 0.016 0.017 0.017 0.019 0.016 0.021 0.014 0.017 0.014 0.015 0.018 0.015 0.017 0.014 0.017 EXR1356 BHP1222 Caramulla Creek 0.169 0.245 0.226 0.250 0.195 0.191 0.008 0.183 0.183 0.180 0.133 0.133 0.258 0.014 0.013 0.014 0.018 0.019 0.016 0.014 0.020 0.014 0.014 0.014 0.013 0.016 0.015 0.015 0.015 0.015 G121 Symphyla 0.159 0.233 0.216 0.246 0.216 0.205 0.177 0.183 0.180 0.186 0.177 0.177 0.252 0.176 0.012 0.013 0.018 0.016 0.017 0.014 0.021 0.014 0.015 0.015 0.013 0.015 0.015 0.014 0.016 0.014 G366 Symphyla sp OP LN9204 0.174 0.217 0.221 0.252 0.211 0.190 0.178 0.162 0.164 0.177 0.171 0.171 0.244 0.180 0.174 0.013 0.017 0.018 0.019 0.015 0.020 0.014 0.015 0.014 0.015 0.014 0.014 0.016 0.015 0.016 G368 Symphyla sp MA LN8697 0.195 0.228 0.220 0.258 0.211 0.202 0.209 0.184 0.188 0.190 0.196 0.196 0.266 0.212 0.185 0.175 0.017 0.017 0.017 0.015 0.018 0.017 0.016 0.015 0.015 0.014 0.015 0.015 0.017 0.015 G42 Symphyla 0.239 0.261 0.216 0.230 0.221 0.260 0.224 0.219 0.212 0.215 0.228 0.228 0.252 0.220 0.241 0.226 0.239 0.015 0.016 0.017 0.017 0.016 0.016 0.014 0.018 0.018 0.016 0.018 0.017 0.019 G43 Symphyla 0.244 0.181 0.252 0.211 0.261 0.256 0.251 0.233 0.225 0.231 0.251 0.251 0.217 0.244 0.235 0.227 0.225 0.240 0.018 0.017 0.021 0.014 0.019 0.018 0.017 0.019 0.017 0.020 0.015 0.020 G44 Symphyla 0.209 0.264 0.208 0.252 0.247 0.218 0.226 0.228 0.221 0.221 0.245 0.245 0.263 0.224 0.207 0.202 0.224 0.187 0.246 0.013 0.015 0.017 0.016 0.016 0.018 0.018 0.016 0.016 0.018 0.016 G45 Symphyla 0.218 0.253 0.200 0.267 0.238 0.236 0.222 0.213 0.206 0.219 0.231 0.231 0.288 0.224 0.215 0.206 0.220 0.182 0.268 0.118 0.015 0.016 0.014 0.015 0.014 0.016 0.015 0.014 0.015 0.015 G46 Symphyla 0.251 0.295 0.217 0.266 0.251 0.240 0.232 0.244 0.248 0.235 0.216 0.216 0.294 0.235 0.242 0.252 0.234 0.215 0.273 0.178 0.158 0.019 0.021 0.021 0.022 0.023 0.022 0.021 0.019 0.021 G494 111138b Symphyla 0.281 0.223 0.273 0.194 0.282 0.282 0.271 0.256 0.253 0.256 0.271 0.271 0.258 0.269 0.265 0.260 0.280 0.240 0.200 0.257 0.275 0.269 0.015 0.014 0.015 0.016 0.014 0.016 0.016 0.017 GH23 Hanseniella Han-PES16403 0.186 0.255 0.219 0.261 0.217 0.189 0.170 0.180 0.179 0.181 0.182 0.182 0.263 0.176 0.178 0.185 0.190 0.224 0.252 0.219 0.227 0.252 0.278 0.012 0.014 0.015 0.015 0.015 0.015 0.015 IS72 KBRC1475 20150708 T2 02 Symphyla 0.150 0.234 0.221 0.252 0.208 0.205 0.170 0.168 0.163 0.179 0.187 0.187 0.250 0.177 0.197 0.196 0.217 0.207 0.257 0.229 0.210 0.241 0.277 0.181 0.015 0.017 0.014 0.015 0.015 0.015 J7-22 Wheelarra Hill 0.173 0.215 0.241 0.258 0.198 0.202 0.191 0.162 0.165 0.170 0.185 0.185 0.241 0.191 0.171 0.169 0.185 0.254 0.238 0.232 0.223 0.274 0.287 0.175 0.197 0.015 0.015 0.016 0.015 0.014 JD04 DD11YOX32020151124-T2-3 Symphyla 0.185 0.235 0.214 0.272 0.231 0.207 0.184 0.201 0.187 0.195 0.189 0.189 0.283 0.182 0.182 0.183 0.195 0.241 0.252 0.217 0.218 0.265 0.293 0.202 0.199 0.178 0.016 0.015 0.015 0.015 JIN0290 BHP779 Hancock Range South 0.208 0.253 0.229 0.276 0.217 0.211 0.200 0.192 0.195 0.226 0.203 0.203 0.276 0.197 0.199 0.186 0.202 0.212 0.262 0.241 0.224 0.269 0.285 0.196 0.197 0.205 0.222 0.014 0.015 0.014 LB069 BHP984 Hashimoto 0.196 0.265 0.199 0.288 0.219 0.029 0.193 0.193 0.192 0.235 0.201 0.201 0.270 0.192 0.209 0.184 0.201 0.258 0.260 0.228 0.240 0.244 0.277 0.194 0.199 0.212 0.218 0.195 0.015 0.006 MUDUNK0 J14-4 S Boundary R sp B4 0.264 0.167 0.255 0.191 0.263 0.246 0.255 0.231 0.224 0.260 0.247 0.247 0.220 0.252 0.242 0.243 0.269 0.260 0.194 0.274 0.277 0.284 0.217 0.272 0.263 0.250 0.254 0.275 0.244 0.015 PI059 BHP935 Hashimoto 0.185 0.256 0.209 0.284 0.214 0.002 0.191 0.195 0.195 0.228 0.194 0.194 0.273 0.189 0.203 0.189 0.200 0.260 0.256 0.218 0.236 0.240 0.282 0.189 0.203 0.200 0.207 0.209 0.028 0.244 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 22. Estimates of Evolutionary Divergence between Sequences

Lineage 120191_Chilopoda_Cryptopidae 132678_Chilopoda_Cryptopidae 93779_Chilopoda_Cryptopidae_sp. IV142 IV231 Mmult Cjap Smult AK10 BQ12 BX11 BX12 BX13 BX8 BX9 CQ2 CQ3 CQ5 CQ6 DF14 EY5 FH15 FH16 G123 G124 G349 IS87 IV139 IV140 IV141 IV143 Gfla Gbon Char Slin Cpic 120191_Chilopoda_Cryptopidae 0.022 0.024 0.025 0.022 0.024 0.023 0.024 0.023 0.021 0.022 0.023 0.025 0.025 0.025 0.024 0.025 0.022 0.024 0.022 0.024 0.022 0.026 0.022 0.021 0.022 0.024 0.022 0.022 0.021 0.024 0.027 0.024 0.024 0.025 0.024 132678_Chilopoda_Cryptopidae 0.235 0.025 0.025 0.023 0.023 0.023 0.027 0.023 0.024 0.022 0.022 0.020 0.020 0.023 0.024 0.020 0.023 0.023 0.024 0.023 0.026 0.030 0.025 0.024 0.024 0.024 0.026 0.026 0.026 0.025 0.026 0.025 0.024 0.025 0.022 93779_Chilopoda_Cryptopidae_sp. 0.256 0.270 0.019 0.019 0.018 0.019 0.018 0.018 0.018 0.021 0.018 0.021 0.021 0.021 0.018 0.018 0.018 0.020 0.019 0.023 0.020 0.022 0.018 0.018 0.019 0.019 0.019 0.019 0.019 0.020 0.019 0.020 0.016 0.018 0.019 IV142_138572_Chilopoda 0.305 0.284 0.211 0.020 0.020 0.019 0.015 0.020 0.019 0.020 0.021 0.021 0.021 0.021 0.019 0.020 0.020 0.021 0.019 0.022 0.019 0.020 0.017 0.018 0.020 0.019 0.018 0.018 0.018 0.019 0.018 0.020 0.019 0.020 0.023 IV231_RC14MEB0115-20160317-T2-02_Chilopoda 0.200 0.232 0.263 0.248 0.018 0.016 0.018 0.018 0.017 0.021 0.020 0.019 0.020 0.018 0.016 0.017 0.014 0.016 0.016 0.020 0.016 0.017 0.020 0.016 0.016 0.019 0.016 0.016 0.016 0.017 0.021 0.019 0.019 0.020 0.018 AB610774_Mecistocephalus_multidentatus 0.227 0.262 0.258 0.254 0.220 0.017 0.016 0.018 0.017 0.020 0.021 0.020 0.020 0.019 0.018 0.018 0.019 0.021 0.018 0.019 0.020 0.021 0.019 0.019 0.018 0.017 0.017 0.017 0.017 0.019 0.016 0.017 0.019 0.021 0.020 AB610777_Cryptops_japonicus 0.205 0.222 0.234 0.244 0.163 0.216 0.019 0.017 0.018 0.019 0.018 0.020 0.020 0.018 0.019 0.016 0.017 0.018 0.018 0.017 0.017 0.020 0.020 0.017 0.018 0.019 0.018 0.018 0.018 0.017 0.021 0.020 0.020 0.020 0.016 AB614405_Scolopendra_multidens 0.270 0.277 0.196 0.183 0.240 0.216 0.234 0.017 0.019 0.021 0.021 0.019 0.019 0.019 0.019 0.018 0.019 0.020 0.017 0.021 0.018 0.018 0.019 0.017 0.018 0.017 0.016 0.016 0.016 0.018 0.017 0.017 0.019 0.020 0.021 AK10_PE111022_Cryptop_DC10 0.224 0.200 0.236 0.270 0.219 0.250 0.192 0.223 0.018 0.016 0.020 0.018 0.018 0.018 0.019 0.017 0.018 0.017 0.017 0.016 0.016 0.018 0.019 0.019 0.017 0.019 0.018 0.018 0.018 0.017 0.020 0.018 0.019 0.020 0.019 BQ12_PES_5087_Cryptops_won1 0.192 0.210 0.244 0.238 0.185 0.226 0.184 0.245 0.198 0.020 0.020 0.021 0.020 0.021 0.017 0.017 0.017 0.018 0.016 0.020 0.017 0.022 0.019 0.017 0.016 0.021 0.018 0.018 0.018 0.018 0.020 0.020 0.020 0.020 0.018 BX11_PES-0968_Cryptops_pilbara1 0.226 0.198 0.281 0.266 0.226 0.240 0.209 0.242 0.194 0.213 0.016 0.017 0.017 0.017 0.019 0.016 0.018 0.017 0.019 0.009 0.020 0.021 0.021 0.020 0.019 0.020 0.019 0.019 0.019 0.019 0.020 0.021 0.020 0.019 0.019 BX12_PES-3783_Cryptops_pilbara2 0.233 0.209 0.263 0.259 0.225 0.229 0.211 0.258 0.219 0.233 0.169 0.016 0.017 0.018 0.019 0.017 0.018 0.016 0.021 0.017 0.020 0.021 0.019 0.019 0.020 0.020 0.020 0.020 0.020 0.017 0.020 0.022 0.020 0.019 0.018 BX13_991-5262_Cryptops_MH1 0.226 0.221 0.270 0.273 0.214 0.246 0.211 0.254 0.218 0.236 0.201 0.184 0.003 0.017 0.020 0.013 0.020 0.016 0.017 0.019 0.021 0.022 0.020 0.021 0.022 0.019 0.020 0.020 0.020 0.019 0.020 0.024 0.022 0.021 0.020 BX8_PES-5889_Cryptops_MH2 0.226 0.221 0.269 0.272 0.214 0.248 0.211 0.252 0.215 0.233 0.201 0.181 0.006 0.016 0.020 0.013 0.020 0.017 0.017 0.019 0.021 0.022 0.020 0.020 0.022 0.018 0.020 0.020 0.020 0.018 0.020 0.024 0.022 0.021 0.019 BX9_PES-5272_Cryptops_MH2 0.225 0.210 0.278 0.269 0.196 0.247 0.199 0.252 0.208 0.218 0.173 0.186 0.175 0.175 0.020 0.015 0.020 0.016 0.019 0.016 0.019 0.022 0.018 0.020 0.020 0.018 0.018 0.018 0.017 0.017 0.021 0.023 0.020 0.019 0.017 CQ2_nr_Cryptops_sp._B15_CCWUNK05 0.219 0.228 0.250 0.235 0.201 0.228 0.199 0.243 0.207 0.192 0.210 0.194 0.228 0.222 0.228 0.018 0.017 0.016 0.017 0.020 0.017 0.020 0.018 0.017 0.018 0.017 0.020 0.020 0.020 0.016 0.018 0.020 0.018 0.019 0.019 CQ3_Cryptops_sp._B32_CC1798 0.236 0.207 0.256 0.265 0.207 0.229 0.195 0.243 0.211 0.198 0.193 0.174 0.127 0.124 0.171 0.197 0.017 0.016 0.015 0.018 0.017 0.021 0.018 0.018 0.018 0.016 0.019 0.019 0.019 0.017 0.019 0.022 0.019 0.019 0.017 CQ5_nr_Cryptops_sp._B11_PCRC088 0.206 0.239 0.222 0.269 0.169 0.236 0.157 0.231 0.204 0.194 0.209 0.218 0.220 0.225 0.229 0.185 0.205 0.018 0.016 0.021 0.016 0.016 0.018 0.014 0.017 0.020 0.019 0.019 0.019 0.016 0.020 0.020 0.018 0.019 0.017 CQ6_Cryptops_sp._B30_1106_13_EX13 0.233 0.214 0.252 0.265 0.209 0.246 0.199 0.245 0.193 0.220 0.178 0.153 0.178 0.175 0.167 0.204 0.164 0.205 0.018 0.018 0.018 0.022 0.018 0.018 0.019 0.021 0.018 0.018 0.018 0.018 0.021 0.024 0.020 0.020 0.018 DF14_9703_Cryptopidae_sp. 0.180 0.246 0.242 0.245 0.178 0.214 0.196 0.243 0.204 0.161 0.225 0.218 0.229 0.229 0.216 0.175 0.190 0.172 0.205 0.019 0.015 0.017 0.017 0.015 0.014 0.018 0.017 0.017 0.017 0.016 0.020 0.019 0.019 0.019 0.018 EY5_Cryptops_sp_B35 0.228 0.200 0.287 0.266 0.225 0.241 0.197 0.241 0.197 0.225 0.062 0.157 0.213 0.210 0.150 0.208 0.180 0.218 0.176 0.224 0.020 0.022 0.022 0.020 0.020 0.021 0.018 0.018 0.018 0.020 0.021 0.021 0.020 0.018 0.019 FH15_nr_Cryptops_sp_B34_DL0414 0.206 0.239 0.253 0.243 0.178 0.236 0.175 0.234 0.180 0.178 0.220 0.231 0.226 0.229 0.224 0.195 0.204 0.164 0.214 0.166 0.222 0.019 0.020 0.016 0.015 0.019 0.018 0.018 0.018 0.017 0.020 0.022 0.020 0.021 0.017 FH16_nr_Cryptops_sp_B34_EW0769 0.202 0.257 0.227 0.243 0.174 0.235 0.190 0.213 0.186 0.192 0.206 0.231 0.228 0.228 0.222 0.187 0.212 0.145 0.237 0.177 0.205 0.168 0.021 0.018 0.018 0.021 0.022 0.022 0.022 0.020 0.020 0.020 0.021 0.021 0.020 G123_Cryptops_Upper_South_Fortescue 0.262 0.256 0.173 0.177 0.241 0.246 0.217 0.189 0.222 0.237 0.242 0.236 0.257 0.254 0.250 0.236 0.248 0.230 0.213 0.214 0.261 0.232 0.219 0.019 0.019 0.021 0.018 0.018 0.018 0.020 0.017 0.017 0.018 0.020 0.021 G124_Cryptops_Upper_South_Fortescue 0.169 0.249 0.249 0.248 0.173 0.236 0.189 0.236 0.220 0.170 0.227 0.236 0.228 0.231 0.225 0.209 0.220 0.182 0.221 0.168 0.225 0.180 0.179 0.241 0.015 0.018 0.017 0.017 0.017 0.018 0.021 0.022 0.020 0.021 0.019 G349_Cryptopidae_sp_MJ_LN9808 0.182 0.242 0.221 0.251 0.183 0.243 0.185 0.235 0.203 0.158 0.220 0.239 0.231 0.228 0.223 0.192 0.195 0.182 0.217 0.165 0.227 0.184 0.158 0.226 0.166 0.018 0.019 0.019 0.020 0.017 0.020 0.019 0.018 0.020 0.018 IS87_KBRC136220150911T2_03_Geophilida_sp 0.252 0.246 0.256 0.242 0.231 0.188 0.219 0.240 0.239 0.225 0.230 0.231 0.248 0.248 0.231 0.225 0.229 0.225 0.244 0.219 0.229 0.208 0.240 0.239 0.236 0.219 0.019 0.019 0.019 0.019 0.018 0.021 0.020 0.020 0.019 IV139_DD14MRR0004T1-01_Chilopoda 0.236 0.258 0.256 0.235 0.229 0.191 0.219 0.211 0.252 0.232 0.250 0.234 0.237 0.239 0.232 0.246 0.227 0.236 0.227 0.225 0.241 0.232 0.239 0.235 0.242 0.240 0.227 0.000 0.002 0.017 0.020 0.020 0.017 0.021 0.020 IV140_DD14MRR0004T1-01_Chilopoda 0.236 0.258 0.256 0.235 0.229 0.191 0.219 0.211 0.252 0.232 0.250 0.234 0.237 0.239 0.232 0.246 0.227 0.236 0.227 0.225 0.241 0.232 0.239 0.235 0.242 0.240 0.227 0.000 0.002 0.017 0.020 0.020 0.017 0.021 0.020 IV141_DD14MRR0005-T1-04_Chilopoda 0.234 0.258 0.255 0.235 0.229 0.190 0.217 0.208 0.251 0.232 0.246 0.233 0.237 0.239 0.229 0.246 0.227 0.233 0.228 0.225 0.238 0.231 0.235 0.232 0.241 0.240 0.225 0.003 0.003 0.017 0.020 0.020 0.017 0.020 0.020 IV143_MEARC4383-20151001-T2-01_Chilopoda 0.215 0.237 0.224 0.258 0.195 0.229 0.175 0.223 0.211 0.215 0.227 0.209 0.230 0.225 0.204 0.198 0.220 0.180 0.220 0.193 0.202 0.186 0.187 0.234 0.204 0.211 0.219 0.230 0.230 0.228 0.021 0.021 0.019 0.018 0.019 JN306685_Geophilus_flavus 0.274 0.262 0.264 0.222 0.264 0.217 0.249 0.234 0.245 0.237 0.243 0.260 0.271 0.272 0.271 0.240 0.246 0.257 0.264 0.254 0.236 0.239 0.260 0.243 0.265 0.245 0.231 0.223 0.223 0.220 0.281 0.019 0.018 0.017 0.020 KF569297_Gnathoribautia_bonensis 0.249 0.261 0.246 0.252 0.223 0.202 0.197 0.226 0.229 0.227 0.241 0.249 0.260 0.262 0.255 0.238 0.246 0.233 0.255 0.220 0.249 0.251 0.211 0.223 0.232 0.224 0.239 0.206 0.206 0.206 0.235 0.194 0.019 0.020 0.023 KF676531_Cormocephalus_hartmeyeri_Scolopendridae 0.277 0.263 0.164 0.195 0.231 0.237 0.219 0.187 0.216 0.239 0.250 0.252 0.261 0.256 0.263 0.229 0.243 0.232 0.228 0.250 0.271 0.239 0.215 0.148 0.244 0.220 0.258 0.229 0.229 0.227 0.234 0.253 0.225 0.018 0.018 KR736251_Stenotaenia_linearis 0.298 0.298 0.275 0.268 0.278 0.240 0.271 0.260 0.272 0.262 0.268 0.260 0.271 0.272 0.266 0.283 0.254 0.272 0.271 0.267 0.268 0.272 0.262 0.267 0.271 0.280 0.264 0.245 0.245 0.243 0.271 0.196 0.216 0.261 0.020 JF273290_Cryptops_pictus 0.229 0.190 0.277 0.302 0.206 0.246 0.211 0.274 0.218 0.217 0.212 0.199 0.210 0.207 0.196 0.209 0.199 0.215 0.196 0.228 0.190 0.214 0.223 0.269 0.225 0.223 0.245 0.234 0.234 0.231 0.221 0.272 0.267 0.256 0.274 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Table 25. Estimates of Evolutionary Divergence between Sequences

Lineage IV232 IV234 IV392 IV395 IV405 IV413 IV415 IV419 IV422 IV423 AX43 AX62 BX6 BX7 CA4 FH1 G202 G203 G216 Eibl Trig-sp Ooris IE15 IE17 IE19 IE25 IE27 IE28 IE30 IV147 IV150 IV151 IV152 IV153 JV01 JV02 JV03 JV04 JV05 Nviol Stob LN6119 WAME89084 WAME89085 WAME89094 WAME89101 WAME89111 WAME89155 WAME89168 WAME89176 WAME89186 WAME89188 G114 G115 G122 G436 G437 Ptil-sp Blug Htar Lser Dfris Aflav KP422158 Onig Cu-sp. 8 IV232 RC15MEB0107-20160317-T2-02 Coleoptera 0.014 0.015 0.017 0.014 0.013 0.014 0.017 0.013 0.015 0.015 0.014 0.019 0.017 0.012 0.015 0.018 0.016 0.015 0.015 0.014 0.013 0.014 0.014 0.014 0.019 0.013 0.013 0.016 0.015 0.014 0.007 0.015 0.016 0.016 0.016 0.014 0.014 0.014 0.015 0.017 0.019 0.016 0.017 0.015 0.015 0.015 0.016 0.015 0.016 0.014 0.015 0.019 0.019 0.017 0.015 0.019 0.015 0.012 0.014 0.014 0.014 0.016 0.015 0.015 0.014 IV234 RC15MEC0192-20160316-T3-03 Coleoptera 0.232 0.017 0.013 0.014 0.014 0.016 0.016 0.015 0.015 0.015 0.016 0.021 0.020 0.016 0.016 0.021 0.020 0.020 0.015 0.014 0.016 0.014 0.014 0.014 0.024 0.014 0.015 0.016 0.015 0.000 0.015 0.007 0.015 0.017 0.018 0.015 0.016 0.015 0.017 0.015 0.025 0.014 0.015 0.014 0.014 0.015 0.015 0.014 0.016 0.006 0.007 0.021 0.020 0.021 0.020 0.022 0.014 0.017 0.017 0.015 0.015 0.015 0.016 0.016 0.006 IV392 DD14MEC0006T2-02 Coleoptera 0.201 0.263 0.017 0.015 0.015 0.016 0.016 0.014 0.016 0.014 0.014 0.020 0.020 0.015 0.016 0.020 0.018 0.020 0.016 0.016 0.015 0.015 0.015 0.018 0.002 0.015 0.016 0.015 0.015 0.017 0.014 0.017 0.015 0.015 0.016 0.012 0.014 0.013 0.014 0.015 0.019 0.014 0.015 0.014 0.013 0.014 0.017 0.015 0.016 0.018 0.018 0.020 0.019 0.019 0.018 0.019 0.015 0.015 0.017 0.014 0.013 0.016 0.013 0.015 0.018 IV395 MEK1478P7T2-2 Coleoptera 0.246 0.129 0.260 0.014 0.016 0.017 0.016 0.016 0.017 0.014 0.015 0.021 0.020 0.018 0.016 0.023 0.021 0.020 0.016 0.015 0.017 0.014 0.014 0.015 0.023 0.015 0.016 0.017 0.016 0.013 0.018 0.013 0.017 0.017 0.017 0.017 0.016 0.017 0.017 0.017 0.022 0.016 0.016 0.016 0.015 0.016 0.017 0.015 0.016 0.013 0.014 0.019 0.020 0.022 0.021 0.021 0.015 0.017 0.016 0.017 0.017 0.015 0.016 0.018 0.013 IV405 MEK1721P7T1-3 Coleoptera 0.204 0.262 0.222 0.262 0.017 0.016 0.016 0.016 0.014 0.014 0.014 0.020 0.019 0.016 0.016 0.021 0.019 0.016 0.016 0.016 0.018 0.007 0.002 0.017 0.018 0.015 0.015 0.017 0.016 0.014 0.014 0.014 0.016 0.016 0.016 0.016 0.014 0.016 0.016 0.016 0.022 0.017 0.016 0.016 0.016 0.016 0.017 0.005 0.015 0.014 0.014 0.022 0.021 0.021 0.020 0.019 0.005 0.015 0.016 0.017 0.016 0.016 0.016 0.016 0.014 IV413 MEGRC0097P6T1-5 Coleoptera 0.226 0.246 0.213 0.256 0.226 0.016 0.015 0.014 0.016 0.015 0.014 0.019 0.020 0.015 0.015 0.019 0.019 0.020 0.016 0.018 0.015 0.016 0.016 0.016 0.021 0.014 0.015 0.013 0.017 0.014 0.014 0.015 0.014 0.013 0.016 0.015 0.016 0.016 0.015 0.014 0.021 0.014 0.015 0.014 0.014 0.015 0.017 0.017 0.016 0.014 0.015 0.024 0.019 0.019 0.018 0.018 0.017 0.014 0.016 0.014 0.014 0.016 0.014 0.014 0.014 IV415 RC08SILV0683p3t1-1 Coleoptera 0.138 0.237 0.203 0.237 0.207 0.225 0.015 0.014 0.015 0.016 0.015 0.019 0.019 0.013 0.015 0.019 0.018 0.016 0.016 0.015 0.014 0.016 0.016 0.015 0.021 0.014 0.013 0.016 0.015 0.016 0.013 0.015 0.015 0.016 0.016 0.015 0.013 0.014 0.014 0.015 0.018 0.017 0.016 0.016 0.014 0.015 0.017 0.017 0.017 0.015 0.016 0.021 0.020 0.018 0.017 0.018 0.017 0.013 0.013 0.015 0.014 0.016 0.014 0.016 0.015 IV419 D08BU075P1T1-4 Coleoptera 0.194 0.249 0.207 0.256 0.207 0.210 0.197 0.016 0.016 0.016 0.014 0.021 0.020 0.016 0.016 0.020 0.020 0.018 0.018 0.016 0.016 0.016 0.017 0.017 0.022 0.016 0.016 0.015 0.017 0.016 0.017 0.016 0.016 0.015 0.016 0.016 0.016 0.017 0.015 0.016 0.021 0.018 0.017 0.017 0.015 0.016 0.017 0.017 0.018 0.016 0.017 0.021 0.022 0.020 0.018 0.020 0.017 0.016 0.016 0.017 0.016 0.016 0.015 0.016 0.016 IV422 E004FP3T1-4 Coleoptera 0.179 0.263 0.186 0.262 0.209 0.223 0.172 0.204 0.014 0.015 0.012 0.019 0.019 0.013 0.016 0.018 0.016 0.018 0.015 0.016 0.014 0.014 0.016 0.015 0.019 0.013 0.013 0.013 0.015 0.015 0.013 0.015 0.015 0.014 0.015 0.014 0.014 0.014 0.013 0.016 0.019 0.015 0.014 0.016 0.015 0.016 0.016 0.015 0.016 0.015 0.016 0.021 0.017 0.017 0.017 0.019 0.015 0.012 0.014 0.002 0.014 0.016 0.013 0.016 0.015 IV423 BUNW01114P1T2-3 Coleoptera 0.186 0.238 0.185 0.243 0.217 0.204 0.190 0.185 0.194 0.015 0.015 0.023 0.022 0.016 0.014 0.023 0.021 0.019 0.015 0.016 0.012 0.014 0.014 0.015 0.020 0.016 0.016 0.016 0.016 0.015 0.014 0.016 0.013 0.015 0.015 0.014 0.016 0.016 0.014 0.013 0.019 0.015 0.014 0.015 0.014 0.015 0.016 0.015 0.017 0.015 0.016 0.020 0.019 0.022 0.021 0.019 0.015 0.014 0.015 0.015 0.014 0.013 0.014 0.015 0.015 AX43 ID29P1T1-3 Coleoptera 0.191 0.248 0.159 0.237 0.221 0.220 0.198 0.215 0.206 0.196 0.015 0.020 0.020 0.014 0.015 0.023 0.019 0.019 0.015 0.016 0.013 0.015 0.014 0.014 0.017 0.015 0.014 0.016 0.018 0.015 0.014 0.014 0.016 0.015 0.015 0.013 0.014 0.014 0.015 0.016 0.022 0.014 0.015 0.013 0.014 0.014 0.017 0.015 0.017 0.015 0.015 0.019 0.019 0.020 0.019 0.019 0.015 0.016 0.016 0.015 0.016 0.017 0.015 0.015 0.015 AX62 11DDH051P1T2-3 Coleoptera 0.179 0.238 0.178 0.234 0.198 0.217 0.176 0.169 0.160 0.180 0.177 0.020 0.019 0.015 0.015 0.019 0.019 0.018 0.017 0.016 0.014 0.014 0.014 0.015 0.019 0.014 0.015 0.016 0.016 0.016 0.015 0.016 0.016 0.016 0.015 0.014 0.014 0.015 0.013 0.017 0.021 0.015 0.015 0.014 0.016 0.015 0.017 0.015 0.017 0.017 0.017 0.019 0.019 0.019 0.018 0.019 0.015 0.014 0.015 0.014 0.016 0.018 0.015 0.015 0.017 BX6 PES-5290 Bembidion MH1 CaK Carabidae 0.194 0.302 0.233 0.300 0.221 0.212 0.205 0.235 0.221 0.224 0.233 0.209 0.007 0.017 0.020 0.017 0.015 0.018 0.021 0.022 0.021 0.021 0.020 0.020 0.021 0.018 0.019 0.020 0.019 0.021 0.020 0.022 0.020 0.020 0.022 0.019 0.018 0.019 0.020 0.020 0.018 0.019 0.020 0.020 0.020 0.020 0.025 0.020 0.021 0.023 0.022 0.022 0.016 0.019 0.016 0.019 0.020 0.018 0.020 0.018 0.019 0.019 0.023 0.020 0.023 BX7 PES-5285 Bembidion MH1 CaK Carabidae 0.177 0.297 0.219 0.295 0.212 0.219 0.194 0.233 0.217 0.233 0.224 0.209 0.031 0.017 0.019 0.018 0.014 0.017 0.020 0.021 0.020 0.020 0.019 0.020 0.021 0.018 0.017 0.021 0.020 0.020 0.018 0.021 0.019 0.020 0.022 0.018 0.018 0.019 0.020 0.019 0.019 0.019 0.019 0.019 0.018 0.018 0.024 0.019 0.021 0.021 0.021 0.022 0.017 0.017 0.016 0.018 0.019 0.018 0.019 0.018 0.019 0.018 0.022 0.020 0.021 CA4 11:1461 Anillini sp. McP1 0.130 0.243 0.186 0.244 0.197 0.226 0.142 0.179 0.178 0.198 0.174 0.165 0.164 0.162 0.016 0.019 0.015 0.018 0.017 0.015 0.014 0.015 0.015 0.017 0.020 0.015 0.014 0.016 0.017 0.016 0.013 0.017 0.015 0.015 0.016 0.011 0.013 0.014 0.014 0.015 0.017 0.016 0.016 0.016 0.015 0.015 0.018 0.016 0.016 0.017 0.017 0.022 0.018 0.016 0.015 0.016 0.016 0.012 0.014 0.014 0.014 0.016 0.017 0.013 0.017 FH1 Curculionidae Genus2 B15 EW0540 0.235 0.234 0.225 0.237 0.247 0.232 0.225 0.232 0.231 0.229 0.221 0.212 0.262 0.255 0.224 0.022 0.020 0.019 0.015 0.016 0.015 0.015 0.016 0.015 0.021 0.014 0.014 0.016 0.016 0.016 0.016 0.016 0.017 0.015 0.014 0.016 0.016 0.015 0.014 0.017 0.022 0.015 0.015 0.016 0.016 0.015 0.014 0.016 0.015 0.016 0.016 0.019 0.019 0.020 0.020 0.022 0.016 0.015 0.015 0.017 0.016 0.015 0.015 0.016 0.016 G202 Anillini 100490 0.177 0.283 0.237 0.293 0.221 0.230 0.189 0.221 0.175 0.237 0.219 0.189 0.135 0.140 0.153 0.261 0.010 0.017 0.021 0.023 0.020 0.021 0.021 0.021 0.019 0.018 0.018 0.019 0.022 0.021 0.017 0.021 0.020 0.020 0.021 0.017 0.019 0.020 0.020 0.020 0.020 0.020 0.021 0.019 0.020 0.019 0.023 0.022 0.021 0.021 0.021 0.022 0.019 0.011 0.010 0.018 0.022 0.019 0.020 0.018 0.020 0.019 0.022 0.019 0.021 G203 Anillini 107877 0.169 0.279 0.207 0.292 0.213 0.211 0.175 0.220 0.173 0.216 0.199 0.182 0.114 0.110 0.134 0.255 0.050 0.016 0.019 0.021 0.019 0.019 0.019 0.020 0.019 0.017 0.017 0.018 0.020 0.020 0.015 0.020 0.020 0.017 0.019 0.016 0.017 0.019 0.018 0.020 0.019 0.019 0.019 0.018 0.018 0.018 0.023 0.020 0.020 0.021 0.021 0.022 0.016 0.010 0.009 0.015 0.020 0.016 0.018 0.016 0.018 0.018 0.020 0.019 0.021 G216 Coleoptera 109656 0.130 0.261 0.211 0.254 0.196 0.252 0.146 0.213 0.207 0.218 0.196 0.189 0.185 0.171 0.163 0.251 0.171 0.157 0.020 0.020 0.018 0.017 0.016 0.017 0.021 0.017 0.015 0.020 0.021 0.020 0.015 0.020 0.018 0.021 0.022 0.022 0.017 0.021 0.020 0.018 0.019 0.021 0.022 0.021 0.021 0.021 0.021 0.017 0.020 0.021 0.021 0.022 0.019 0.017 0.017 0.017 0.017 0.019 0.017 0.018 0.018 0.018 0.021 0.019 0.021 GU213687 Echinodera ibleiensis Curculionidae 0.228 0.190 0.236 0.213 0.255 0.251 0.229 0.245 0.220 0.229 0.227 0.233 0.284 0.284 0.234 0.219 0.257 0.265 0.244 0.015 0.015 0.016 0.016 0.016 0.021 0.015 0.016 0.018 0.016 0.015 0.016 0.015 0.014 0.017 0.016 0.015 0.015 0.017 0.014 0.014 0.022 0.017 0.017 0.017 0.017 0.018 0.014 0.015 0.016 0.015 0.015 0.020 0.017 0.021 0.020 0.022 0.015 0.015 0.016 0.016 0.015 0.013 0.017 0.018 0.015 HE615891 Trigonopterus sp. Curculionidae 0.215 0.196 0.237 0.224 0.236 0.254 0.224 0.239 0.224 0.225 0.235 0.245 0.298 0.284 0.239 0.205 0.283 0.274 0.260 0.198 0.017 0.016 0.016 0.017 0.023 0.015 0.015 0.017 0.018 0.014 0.015 0.014 0.016 0.017 0.016 0.017 0.016 0.016 0.015 0.016 0.021 0.015 0.015 0.015 0.015 0.015 0.015 0.016 0.015 0.015 0.014 0.018 0.021 0.021 0.021 0.021 0.016 0.014 0.015 0.016 0.016 0.015 0.015 0.016 0.015 HQ953104 Orthospila orissusalis Crambidae 0.191 0.260 0.181 0.272 0.202 0.217 0.179 0.210 0.187 0.187 0.194 0.170 0.222 0.213 0.170 0.214 0.218 0.194 0.187 0.239 0.237 0.017 0.018 0.016 0.019 0.014 0.014 0.015 0.014 0.016 0.013 0.016 0.017 0.015 0.015 0.015 0.014 0.015 0.014 0.016 0.021 0.014 0.015 0.014 0.015 0.015 0.017 0.018 0.016 0.016 0.016 0.020 0.020 0.018 0.018 0.020 0.018 0.013 0.013 0.015 0.014 0.015 0.015 0.014 0.016 IE15 BHRC122-20141216-T2-01 0.201 0.257 0.226 0.266 0.030 0.229 0.203 0.204 0.200 0.219 0.226 0.192 0.225 0.216 0.196 0.246 0.221 0.213 0.204 0.251 0.244 0.196 0.007 0.017 0.018 0.015 0.015 0.016 0.015 0.014 0.015 0.014 0.016 0.016 0.017 0.016 0.014 0.016 0.015 0.016 0.022 0.016 0.017 0.016 0.015 0.016 0.017 0.007 0.014 0.015 0.015 0.022 0.021 0.020 0.020 0.019 0.007 0.015 0.016 0.015 0.016 0.015 0.016 0.016 0.015 IE17 BHRC058-20141215-T1-01 0.203 0.260 0.220 0.260 0.001 0.228 0.206 0.206 0.207 0.216 0.217 0.195 0.220 0.211 0.196 0.245 0.221 0.215 0.197 0.254 0.234 0.201 0.028 0.017 0.018 0.015 0.015 0.017 0.016 0.014 0.014 0.013 0.015 0.016 0.016 0.016 0.014 0.016 0.016 0.016 0.022 0.017 0.016 0.016 0.016 0.016 0.017 0.005 0.015 0.014 0.014 0.022 0.021 0.021 0.020 0.019 0.005 0.014 0.016 0.017 0.016 0.016 0.016 0.016 0.014 IE19 BHRC218-20141216-T2-01 0.180 0.241 0.209 0.231 0.241 0.211 0.200 0.209 0.195 0.170 0.203 0.190 0.229 0.232 0.212 0.229 0.213 0.208 0.213 0.224 0.216 0.207 0.240 0.239 0.021 0.015 0.016 0.017 0.016 0.014 0.014 0.015 0.014 0.016 0.016 0.015 0.018 0.014 0.015 0.015 0.021 0.016 0.015 0.016 0.016 0.017 0.017 0.017 0.017 0.015 0.015 0.019 0.015 0.021 0.019 0.020 0.017 0.016 0.017 0.015 0.014 0.015 0.017 0.016 0.015 IE25 CBRC122P3T2-3 0.214 0.306 0.002 0.301 0.234 0.224 0.210 0.248 0.205 0.207 0.165 0.200 0.241 0.227 0.190 0.243 0.236 0.219 0.219 0.248 0.253 0.195 0.236 0.234 0.236 0.021 0.020 0.020 0.019 0.024 0.018 0.024 0.020 0.019 0.020 0.018 0.018 0.017 0.016 0.019 0.018 0.017 0.017 0.017 0.016 0.017 0.022 0.018 0.020 0.024 0.024 0.021 0.020 0.020 0.019 0.019 0.018 0.019 0.021 0.019 0.018 0.020 0.016 0.020 0.024 IE27 CWRC179P4T3-5 0.133 0.259 0.210 0.256 0.214 0.212 0.156 0.238 0.173 0.197 0.213 0.172 0.151 0.151 0.160 0.238 0.173 0.145 0.179 0.245 0.230 0.190 0.213 0.214 0.184 0.236 0.007 0.016 0.017 0.014 0.012 0.015 0.016 0.016 0.016 0.012 0.015 0.014 0.015 0.016 0.017 0.015 0.016 0.015 0.015 0.014 0.017 0.015 0.016 0.014 0.015 0.023 0.017 0.017 0.017 0.018 0.015 0.013 0.014 0.013 0.015 0.016 0.014 0.014 0.014 IE28 CWRC274P4T3-5 0.135 0.266 0.202 0.261 0.208 0.207 0.154 0.229 0.174 0.190 0.201 0.174 0.152 0.142 0.152 0.236 0.176 0.143 0.162 0.243 0.233 0.186 0.208 0.207 0.192 0.223 0.037 0.017 0.018 0.015 0.011 0.016 0.016 0.016 0.018 0.011 0.016 0.015 0.015 0.017 0.017 0.016 0.016 0.015 0.014 0.014 0.017 0.015 0.017 0.015 0.016 0.023 0.019 0.018 0.017 0.019 0.015 0.012 0.013 0.013 0.015 0.016 0.014 0.015 0.015 IE30 MEBRC0021-0712-T1-2 0.217 0.281 0.223 0.293 0.244 0.182 0.213 0.203 0.207 0.197 0.233 0.218 0.234 0.220 0.205 0.239 0.235 0.217 0.264 0.255 0.271 0.220 0.234 0.242 0.223 0.239 0.226 0.233 0.017 0.016 0.015 0.017 0.014 0.006 0.009 0.015 0.016 0.015 0.016 0.014 0.019 0.015 0.014 0.015 0.013 0.014 0.015 0.017 0.017 0.017 0.018 0.020 0.021 0.018 0.016 0.019 0.017 0.016 0.017 0.013 0.015 0.016 0.015 0.014 0.017 IV147 DD14MRR0004T1-03 Coleoptera 0.206 0.260 0.204 0.265 0.161 0.226 0.203 0.209 0.216 0.201 0.238 0.201 0.222 0.225 0.212 0.236 0.223 0.217 0.224 0.252 0.250 0.166 0.163 0.161 0.226 0.207 0.226 0.235 0.240 0.015 0.016 0.015 0.017 0.015 0.017 0.016 0.015 0.015 0.015 0.017 0.021 0.016 0.017 0.017 0.017 0.017 0.016 0.016 0.015 0.016 0.016 0.020 0.019 0.022 0.019 0.021 0.016 0.015 0.016 0.016 0.014 0.015 0.015 0.019 0.016 IV150 RC14MEB0060-20151001-T2-02 Coleoptera 0.232 0.000 0.263 0.129 0.262 0.246 0.237 0.249 0.263 0.238 0.245 0.236 0.300 0.296 0.240 0.231 0.283 0.280 0.262 0.190 0.196 0.260 0.255 0.258 0.240 0.306 0.260 0.266 0.278 0.258 0.015 0.007 0.015 0.017 0.018 0.015 0.016 0.015 0.017 0.015 0.025 0.014 0.015 0.014 0.014 0.015 0.015 0.014 0.016 0.006 0.007 0.021 0.020 0.021 0.020 0.022 0.014 0.017 0.017 0.015 0.015 0.015 0.016 0.016 0.006 IV151 MEARC3814-20150807-T1-02 Coleoptera 0.037 0.237 0.186 0.235 0.195 0.210 0.124 0.197 0.179 0.185 0.172 0.176 0.197 0.186 0.119 0.225 0.177 0.163 0.141 0.223 0.215 0.179 0.190 0.192 0.185 0.200 0.130 0.120 0.211 0.198 0.234 0.016 0.016 0.015 0.016 0.014 0.015 0.014 0.015 0.016 0.017 0.016 0.016 0.015 0.015 0.014 0.017 0.015 0.016 0.015 0.016 0.021 0.019 0.017 0.014 0.018 0.015 0.012 0.014 0.014 0.014 0.016 0.015 0.014 0.015 IV152 MEARC4400-t1-01 Coleoptera 0.229 0.033 0.269 0.135 0.266 0.256 0.244 0.251 0.274 0.238 0.248 0.234 0.310 0.303 0.242 0.234 0.290 0.286 0.253 0.195 0.204 0.263 0.259 0.262 0.248 0.308 0.267 0.274 0.283 0.271 0.032 0.234 0.015 0.017 0.017 0.015 0.017 0.015 0.016 0.015 0.025 0.015 0.015 0.015 0.015 0.015 0.014 0.014 0.016 0.005 0.001 0.021 0.020 0.020 0.020 0.022 0.014 0.017 0.016 0.016 0.016 0.015 0.016 0.016 0.005 IV153 MEARC4400-t1-01 Coleoptera 0.192 0.241 0.200 0.244 0.216 0.201 0.191 0.194 0.185 0.146 0.216 0.184 0.222 0.216 0.196 0.234 0.209 0.192 0.208 0.222 0.216 0.222 0.217 0.212 0.185 0.222 0.195 0.204 0.206 0.211 0.239 0.198 0.236 0.014 0.015 0.016 0.015 0.016 0.014 0.004 0.019 0.014 0.014 0.013 0.013 0.013 0.017 0.016 0.017 0.015 0.015 0.020 0.021 0.020 0.019 0.019 0.016 0.015 0.015 0.016 0.014 0.013 0.016 0.015 0.015 JV01 GR14BS40032-20160608-T1-01 Coleoptera 0.216 0.283 0.226 0.293 0.249 0.185 0.212 0.210 0.210 0.194 0.235 0.219 0.226 0.224 0.192 0.243 0.235 0.216 0.265 0.254 0.269 0.228 0.240 0.246 0.220 0.243 0.224 0.235 0.026 0.239 0.280 0.208 0.284 0.212 0.008 0.014 0.015 0.015 0.015 0.014 0.019 0.014 0.013 0.015 0.014 0.014 0.016 0.016 0.018 0.017 0.017 0.021 0.019 0.019 0.017 0.019 0.016 0.015 0.016 0.014 0.014 0.016 0.015 0.015 0.017 JV02 RC11BS3045-20160609-T1-01 Coleoptera 0.212 0.283 0.215 0.284 0.240 0.201 0.209 0.202 0.204 0.186 0.222 0.206 0.224 0.227 0.187 0.236 0.233 0.219 0.250 0.257 0.249 0.207 0.241 0.239 0.211 0.231 0.203 0.219 0.060 0.233 0.282 0.205 0.276 0.198 0.058 0.015 0.017 0.016 0.015 0.015 0.018 0.015 0.015 0.015 0.017 0.016 0.016 0.017 0.016 0.017 0.017 0.019 0.019 0.020 0.018 0.019 0.017 0.015 0.016 0.015 0.015 0.015 0.017 0.016 0.017 JV03 RC15BS40051-20160608-T3-01A Coleoptera 0.173 0.231 0.182 0.238 0.192 0.197 0.144 0.192 0.155 0.209 0.189 0.156 0.183 0.174 0.128 0.222 0.170 0.160 0.183 0.217 0.231 0.152 0.186 0.189 0.215 0.186 0.163 0.163 0.211 0.193 0.228 0.160 0.236 0.204 0.204 0.198 0.015 0.014 0.013 0.016 0.018 0.015 0.015 0.015 0.016 0.016 0.018 0.016 0.017 0.015 0.016 0.022 0.018 0.017 0.016 0.017 0.016 0.011 0.013 0.015 0.014 0.015 0.014 0.014 0.015 JV04 RC15BS40051-20160608-T3-01B Coleoptera 0.201 0.262 0.148 0.265 0.204 0.206 0.170 0.220 0.186 0.182 0.171 0.191 0.200 0.196 0.201 0.236 0.223 0.193 0.206 0.239 0.231 0.195 0.204 0.201 0.212 0.152 0.194 0.199 0.217 0.193 0.259 0.192 0.266 0.184 0.222 0.214 0.194 0.014 0.014 0.015 0.022 0.016 0.014 0.015 0.015 0.015 0.017 0.014 0.016 0.017 0.017 0.018 0.020 0.018 0.017 0.018 0.014 0.014 0.013 0.014 0.014 0.016 0.014 0.016 0.017 JV05 RC15BS4B0415-20160609-T2-01 Coleoptera 0.194 0.225 0.149 0.241 0.223 0.213 0.170 0.209 0.198 0.210 0.170 0.179 0.219 0.221 0.175 0.220 0.228 0.189 0.183 0.249 0.231 0.167 0.221 0.220 0.198 0.171 0.198 0.191 0.242 0.214 0.223 0.176 0.225 0.217 0.241 0.220 0.171 0.172 0.013 0.016 0.020 0.015 0.016 0.014 0.015 0.016 0.017 0.015 0.016 0.016 0.016 0.023 0.018 0.020 0.019 0.022 0.015 0.015 0.014 0.014 0.013 0.016 0.013 0.015 0.016 KJ961916 Necrobia violacea voucher ZMUO Cleridae 0.184 0.240 0.163 0.251 0.193 0.182 0.188 0.169 0.172 0.150 0.209 0.147 0.213 0.203 0.173 0.202 0.197 0.175 0.217 0.216 0.218 0.153 0.184 0.191 0.194 0.161 0.179 0.176 0.205 0.166 0.240 0.175 0.243 0.170 0.213 0.191 0.149 0.179 0.175 0.014 0.021 0.014 0.015 0.014 0.013 0.014 0.016 0.016 0.016 0.016 0.016 0.022 0.018 0.019 0.017 0.019 0.016 0.014 0.014 0.013 0.014 0.014 0.014 0.015 0.016 KJ962132 Stricticollis tobias Anthicidae 0.198 0.249 0.204 0.254 0.225 0.204 0.196 0.198 0.191 0.146 0.221 0.190 0.225 0.222 0.202 0.243 0.211 0.194 0.214 0.223 0.218 0.220 0.228 0.223 0.188 0.219 0.193 0.204 0.210 0.219 0.249 0.205 0.246 0.009 0.216 0.196 0.210 0.190 0.222 0.166 0.019 0.014 0.014 0.014 0.014 0.014 0.016 0.016 0.016 0.015 0.015 0.020 0.021 0.021 0.019 0.019 0.016 0.015 0.014 0.016 0.014 0.014 0.016 0.016 0.015 LN6119 CA0021 0.181 0.285 0.230 0.290 0.241 0.228 0.179 0.235 0.228 0.219 0.255 0.228 0.180 0.180 0.194 0.282 0.185 0.173 0.200 0.280 0.278 0.221 0.240 0.244 0.236 0.223 0.185 0.200 0.253 0.240 0.289 0.181 0.284 0.227 0.256 0.231 0.203 0.239 0.211 0.194 0.221 0.020 0.021 0.021 0.022 0.022 0.024 0.022 0.020 0.025 0.025 0.022 0.019 0.020 0.018 0.018 0.022 0.018 0.020 0.019 0.019 0.019 0.023 0.020 0.025 WAME89084 0.225 0.248 0.181 0.258 0.245 0.237 0.207 0.240 0.217 0.223 0.175 0.205 0.236 0.234 0.219 0.217 0.237 0.242 0.219 0.236 0.231 0.191 0.243 0.243 0.218 0.164 0.231 0.232 0.233 0.217 0.248 0.222 0.251 0.222 0.236 0.228 0.208 0.187 0.184 0.205 0.220 0.248 0.007 0.004 0.010 0.010 0.016 0.015 0.016 0.015 0.015 0.022 0.019 0.020 0.019 0.020 0.015 0.015 0.016 0.015 0.014 0.015 0.016 0.015 0.015 WAME89085 0.220 0.254 0.179 0.264 0.248 0.245 0.205 0.233 0.214 0.222 0.168 0.210 0.234 0.222 0.213 0.222 0.235 0.233 0.221 0.240 0.236 0.196 0.243 0.246 0.218 0.159 0.237 0.237 0.223 0.225 0.254 0.219 0.257 0.226 0.225 0.225 0.196 0.179 0.187 0.202 0.225 0.251 0.043 0.008 0.010 0.010 0.017 0.016 0.016 0.015 0.015 0.022 0.020 0.020 0.019 0.020 0.016 0.015 0.015 0.014 0.014 0.015 0.015 0.015 0.015 WAME89094 0.220 0.246 0.179 0.255 0.237 0.236 0.199 0.239 0.217 0.222 0.168 0.199 0.234 0.232 0.214 0.223 0.233 0.237 0.212 0.240 0.230 0.188 0.239 0.236 0.213 0.166 0.222 0.223 0.231 0.222 0.246 0.213 0.249 0.225 0.236 0.227 0.204 0.182 0.181 0.201 0.223 0.246 0.012 0.050 0.010 0.010 0.016 0.016 0.016 0.015 0.015 0.022 0.020 0.019 0.018 0.019 0.016 0.015 0.016 0.016 0.015 0.015 0.015 0.014 0.015 WAME89101 0.211 0.248 0.161 0.251 0.252 0.229 0.193 0.216 0.208 0.210 0.157 0.204 0.248 0.251 0.210 0.223 0.233 0.221 0.205 0.239 0.234 0.207 0.248 0.251 0.210 0.145 0.222 0.218 0.225 0.228 0.248 0.204 0.249 0.225 0.226 0.223 0.211 0.182 0.184 0.196 0.223 0.246 0.087 0.087 0.087 0.006 0.015 0.015 0.016 0.014 0.015 0.023 0.018 0.018 0.017 0.018 0.015 0.016 0.015 0.015 0.014 0.014 0.013 0.014 0.014 WAME89111 0.217 0.249 0.169 0.249 0.252 0.236 0.190 0.214 0.208 0.210 0.157 0.199 0.251 0.248 0.214 0.219 0.235 0.228 0.210 0.246 0.234 0.204 0.251 0.251 0.212 0.152 0.226 0.223 0.228 0.219 0.249 0.208 0.251 0.223 0.233 0.223 0.205 0.172 0.187 0.190 0.222 0.235 0.085 0.084 0.082 0.029 0.016 0.016 0.016 0.015 0.015 0.023 0.019 0.018 0.017 0.019 0.016 0.016 0.015 0.016 0.014 0.014 0.013 0.014 0.015 WAME89155 0.261 0.242 0.249 0.245 0.274 0.278 0.254 0.258 0.249 0.243 0.262 0.251 0.296 0.293 0.248 0.188 0.276 0.300 0.290 0.219 0.224 0.249 0.266 0.272 0.244 0.272 0.286 0.293 0.271 0.248 0.242 0.260 0.242 0.260 0.269 0.264 0.257 0.264 0.260 0.229 0.257 0.314 0.267 0.269 0.271 0.258 0.266 0.018 0.017 0.014 0.014 0.020 0.021 0.022 0.022 0.022 0.018 0.018 0.018 0.016 0.017 0.016 0.015 0.017 0.014 WAME89168 0.213 0.272 0.231 0.272 0.012 0.237 0.217 0.219 0.213 0.225 0.232 0.205 0.222 0.217 0.204 0.254 0.223 0.219 0.205 0.251 0.242 0.202 0.030 0.014 0.244 0.234 0.216 0.211 0.251 0.166 0.272 0.204 0.277 0.229 0.249 0.246 0.199 0.214 0.231 0.195 0.233 0.244 0.242 0.243 0.240 0.254 0.257 0.271 0.014 0.014 0.014 0.022 0.021 0.022 0.022 0.020 0.000 0.015 0.016 0.016 0.016 0.016 0.016 0.016 0.014 WAME89176 0.245 0.242 0.271 0.234 0.251 0.263 0.239 0.246 0.260 0.240 0.274 0.269 0.291 0.281 0.263 0.229 0.266 0.270 0.251 0.208 0.215 0.257 0.245 0.249 0.237 0.284 0.258 0.253 0.275 0.237 0.242 0.236 0.237 0.249 0.281 0.268 0.248 0.243 0.252 0.243 0.246 0.253 0.257 0.267 0.252 0.260 0.255 0.240 0.251 0.016 0.016 0.020 0.019 0.020 0.020 0.020 0.014 0.016 0.017 0.016 0.015 0.016 0.019 0.019 0.016 WAME89186 Curculionidae sp 8 0.240 0.029 0.274 0.140 0.274 0.255 0.249 0.251 0.277 0.246 0.258 0.243 0.319 0.314 0.252 0.242 0.281 0.286 0.263 0.193 0.207 0.260 0.272 0.272 0.253 0.308 0.264 0.268 0.290 0.277 0.029 0.245 0.021 0.252 0.289 0.285 0.240 0.275 0.234 0.242 0.251 0.289 0.255 0.260 0.254 0.248 0.249 0.240 0.277 0.237 0.005 0.021 0.020 0.020 0.020 0.022 0.014 0.018 0.016 0.015 0.016 0.014 0.016 0.015 0.000 WAME89188 0.237 0.032 0.277 0.137 0.275 0.264 0.249 0.257 0.280 0.246 0.259 0.245 0.317 0.312 0.252 0.242 0.288 0.293 0.260 0.193 0.204 0.263 0.271 0.274 0.252 0.308 0.267 0.275 0.293 0.280 0.032 0.245 0.002 0.246 0.295 0.281 0.245 0.278 0.236 0.245 0.248 0.289 0.249 0.257 0.248 0.249 0.251 0.240 0.278 0.239 0.021 0.020 0.021 0.020 0.020 0.022 0.014 0.018 0.016 0.016 0.016 0.015 0.016 0.016 0.005 G114 Curculionidae 0.284 0.251 0.263 0.276 0.261 0.284 0.251 0.281 0.284 0.243 0.262 0.243 0.284 0.291 0.273 0.205 0.296 0.299 0.286 0.223 0.235 0.246 0.261 0.261 0.258 0.265 0.291 0.294 0.289 0.261 0.251 0.284 0.261 0.253 0.289 0.276 0.278 0.258 0.266 0.266 0.246 0.253 0.266 0.266 0.266 0.284 0.273 0.225 0.266 0.246 0.261 0.258 0.019 0.023 0.023 0.023 0.022 0.023 0.021 0.021 0.020 0.021 0.023 0.021 0.021 G115 Curculionidae 0.205 0.221 0.228 0.242 0.242 0.219 0.203 0.226 0.192 0.217 0.239 0.221 0.216 0.222 0.219 0.237 0.216 0.195 0.215 0.189 0.208 0.236 0.240 0.243 0.220 0.243 0.204 0.215 0.252 0.234 0.222 0.204 0.227 0.209 0.255 0.242 0.217 0.220 0.215 0.215 0.212 0.230 0.242 0.245 0.245 0.236 0.240 0.240 0.247 0.215 0.226 0.229 0.251 0.018 0.017 0.017 0.021 0.017 0.019 0.017 0.018 0.018 0.021 0.019 0.020 G122 Anillini 0.172 0.267 0.215 0.292 0.224 0.208 0.186 0.219 0.181 0.229 0.218 0.201 0.150 0.145 0.151 0.251 0.058 0.047 0.176 0.259 0.264 0.199 0.219 0.226 0.217 0.227 0.153 0.153 0.221 0.221 0.269 0.172 0.280 0.208 0.230 0.235 0.169 0.208 0.205 0.176 0.208 0.183 0.243 0.231 0.238 0.218 0.225 0.280 0.234 0.264 0.273 0.285 0.311 0.212 0.011 0.018 0.022 0.018 0.018 0.017 0.019 0.018 0.020 0.019 0.020 G436 110879 Anillini sp 0.171 0.271 0.212 0.286 0.222 0.212 0.171 0.214 0.169 0.218 0.203 0.183 0.127 0.127 0.142 0.246 0.041 0.044 0.168 0.249 0.269 0.203 0.221 0.221 0.204 0.224 0.162 0.167 0.213 0.211 0.270 0.172 0.272 0.198 0.216 0.217 0.157 0.204 0.197 0.181 0.201 0.193 0.240 0.229 0.236 0.218 0.216 0.269 0.231 0.251 0.275 0.277 0.296 0.202 0.061 0.016 0.022 0.017 0.017 0.017 0.018 0.018 0.020 0.018 0.020 G437 110722 Anillini sp long 0.182 0.271 0.190 0.282 0.231 0.214 0.182 0.212 0.203 0.212 0.200 0.184 0.167 0.161 0.146 0.246 0.163 0.146 0.161 0.277 0.273 0.207 0.219 0.230 0.230 0.198 0.166 0.178 0.240 0.230 0.270 0.160 0.274 0.228 0.231 0.212 0.163 0.209 0.199 0.181 0.236 0.186 0.229 0.221 0.229 0.197 0.203 0.297 0.231 0.284 0.277 0.282 0.278 0.218 0.165 0.153 0.020 0.018 0.019 0.018 0.020 0.020 0.021 0.018 0.022 KU519734 Ptiliidae sp. 1 Staphyliniforma 0.213 0.272 0.231 0.272 0.012 0.237 0.217 0.219 0.213 0.225 0.232 0.205 0.222 0.217 0.204 0.254 0.223 0.219 0.205 0.251 0.242 0.202 0.030 0.014 0.244 0.234 0.216 0.211 0.251 0.166 0.272 0.204 0.277 0.229 0.249 0.246 0.199 0.214 0.231 0.195 0.233 0.244 0.242 0.243 0.240 0.254 0.257 0.271 0.000 0.251 0.277 0.278 0.266 0.247 0.234 0.231 0.231 0.015 0.016 0.016 0.016 0.016 0.016 0.016 0.014 JN171062 Bembidion lugubre Carabidae 0.133 0.242 0.188 0.253 0.190 0.209 0.148 0.187 0.154 0.172 0.191 0.158 0.203 0.207 0.147 0.221 0.189 0.199 0.195 0.236 0.230 0.157 0.186 0.189 0.193 0.212 0.146 0.150 0.215 0.195 0.243 0.133 0.246 0.194 0.208 0.193 0.134 0.194 0.180 0.167 0.199 0.204 0.188 0.181 0.187 0.184 0.184 0.252 0.195 0.258 0.248 0.251 0.263 0.245 0.192 0.189 0.180 0.195 0.012 0.013 0.013 0.014 0.014 0.016 0.018 KM447718 Harpalus tardus Carabidae 0.163 0.263 0.178 0.278 0.202 0.219 0.116 0.181 0.169 0.175 0.203 0.155 0.184 0.177 0.141 0.233 0.189 0.166 0.184 0.229 0.231 0.167 0.195 0.201 0.199 0.173 0.155 0.147 0.201 0.195 0.263 0.153 0.266 0.188 0.201 0.191 0.141 0.164 0.185 0.144 0.187 0.192 0.208 0.195 0.201 0.196 0.196 0.260 0.205 0.239 0.267 0.266 0.248 0.238 0.178 0.177 0.190 0.205 0.150 0.014 0.014 0.017 0.015 0.014 0.016 KJ680546 Lasioderma serricorne Aniobidae 0.182 0.269 0.185 0.267 0.213 0.224 0.175 0.205 0.002 0.194 0.206 0.164 0.227 0.222 0.179 0.242 0.175 0.182 0.215 0.224 0.224 0.188 0.203 0.211 0.193 0.205 0.167 0.175 0.206 0.220 0.268 0.183 0.279 0.189 0.210 0.201 0.159 0.195 0.196 0.174 0.196 0.232 0.212 0.209 0.212 0.201 0.201 0.258 0.216 0.259 0.281 0.284 0.284 0.200 0.181 0.168 0.207 0.216 0.161 0.167 0.014 0.017 0.013 0.015 0.015 KM578824 Dermestes frischii Dermestidae 0.155 0.235 0.181 0.250 0.204 0.198 0.170 0.156 0.174 0.166 0.199 0.175 0.204 0.206 0.166 0.215 0.204 0.181 0.206 0.220 0.201 0.178 0.203 0.202 0.177 0.200 0.185 0.182 0.214 0.193 0.234 0.151 0.236 0.162 0.211 0.208 0.172 0.185 0.181 0.150 0.166 0.204 0.217 0.201 0.217 0.207 0.210 0.252 0.210 0.242 0.243 0.243 0.263 0.195 0.182 0.187 0.209 0.210 0.170 0.152 0.173 0.015 0.015 0.016 0.016 KM439906 Anthicus flavipes Anthicidae 0.181 0.231 0.207 0.240 0.210 0.228 0.188 0.175 0.184 0.128 0.215 0.184 0.225 0.222 0.187 0.231 0.199 0.198 0.210 0.199 0.225 0.187 0.211 0.208 0.179 0.229 0.208 0.204 0.210 0.201 0.231 0.184 0.228 0.144 0.219 0.196 0.182 0.211 0.204 0.160 0.144 0.205 0.213 0.210 0.213 0.211 0.204 0.243 0.208 0.229 0.229 0.229 0.261 0.196 0.208 0.192 0.218 0.208 0.176 0.191 0.183 0.169 0.017 0.016 0.014 KP422158 Pselaphinae sp. NZAC Staphylinidae 0.214 0.248 0.132 0.269 0.246 0.229 0.185 0.220 0.195 0.214 0.163 0.185 0.241 0.234 0.211 0.245 0.233 0.210 0.217 0.237 0.230 0.204 0.240 0.245 0.224 0.128 0.210 0.204 0.237 0.228 0.248 0.208 0.254 0.220 0.239 0.232 0.182 0.160 0.178 0.179 0.219 0.235 0.202 0.188 0.196 0.175 0.172 0.255 0.249 0.277 0.252 0.254 0.289 0.247 0.213 0.210 0.229 0.249 0.190 0.181 0.197 0.210 0.220 0.014 0.016 KM439290 Orthoperus nigrescens Corylophidae 0.193 0.266 0.202 0.251 0.242 0.163 0.198 0.216 0.207 0.187 0.200 0.177 0.223 0.223 0.183 0.240 0.207 0.199 0.206 0.260 0.250 0.193 0.237 0.240 0.201 0.210 0.186 0.184 0.187 0.248 0.266 0.189 0.271 0.207 0.187 0.195 0.195 0.204 0.186 0.186 0.205 0.219 0.234 0.227 0.228 0.213 0.219 0.262 0.245 0.253 0.268 0.271 0.256 0.225 0.209 0.199 0.199 0.245 0.198 0.186 0.203 0.204 0.218 0.216 0.015 KU519717 Curculionidae sp. 8 0.240 0.029 0.274 0.140 0.274 0.255 0.249 0.251 0.277 0.246 0.258 0.243 0.319 0.314 0.252 0.242 0.281 0.286 0.263 0.193 0.207 0.260 0.272 0.272 0.253 0.308 0.264 0.268 0.290 0.277 0.029 0.245 0.021 0.252 0.289 0.285 0.240 0.275 0.234 0.242 0.251 0.289 0.255 0.260 0.254 0.248 0.249 0.240 0.277 0.237 0.000 0.021 0.261 0.226 0.273 0.275 0.277 0.277 0.248 0.267 0.281 0.243 0.229 0.252 0.268 1. Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. The number of base differences per site from between sequences are shown. Standard error estimate(s) are shown above the diagonal. The analysis involved 42 nucleotide sequences. Codon positions included were 1st+2nd+3rd+Noncoding. All ambiguous positions were removed for each sequence pair. There were a total of 495 positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [1]. Disclaimer: Although utmost care has been taken to ensure the correctness of the caption, the caption text is provided "as is" without any warranty of any kind. Authors advise the user to carefully check the caption prior to its use for any purpose and report any errors or problems to the authors immediately (www.megasoftware.net). In no event shall the authors and their employers be liable for any damages, including but not limited to special, consequential, or other damages. Authors specifically disclaim all other warranties expressed or implied, including but not limited to the determination of suitability of this caption text for a specific purpose, use, or application. Warramboo Troglofauna Assessment

Appendix 5

WAM Short-Range Endemic Categories Summary

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

This page has been left blank intentionally

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Report by Western Australian Museum (WAMTS403)

APPENDIX 1. WAM SHORT-RANGE ENDEMIC CATEGORIES

Taxonomic Certainty Taxonomic Uncertainty Distribution < 10 000km2 Confirmed SRE Potential SRE • A known distribution of < 10 000km2. • Patchy sampling has resulted in • The taxonomy is well known. incomplete knowledge of the • The group is well represented in geographic distribution of the group. collections and/ or via • We have incomplete taxonomic comprehensive sampling. knowledge. • The group is not well represented in

2 collections. Distribution > 10 000km Widespread (not an SRE) • This category is most applicable to • A known distribution of > 10 000km2. situations where there are gaps in • The taxonomy is well known. our knowledge of the taxon. • The group is well represented in collections and/ or via Sub-categories for this SRE designation comprehensive sampling. are outlined below

SRE SUB-CATEGORIES If a taxon is determined to be a “Potential SRE”, the following sub-categories will further elucidate this status.

A. Data Deficient: • There is insufficient data available to determine SRE status. • Factors that fall under this category include: - New species. - Lack of geographic information. - Lack of taxonomic information. - The group may be poorly represented in collections. - The individuals sampled (e.g. juveniles) may prevent identification to species level.

B. Habitat Indicators: • It is becoming increasingly clear that habitat data can elucidate SRE status. • Where habitat is known to be associated with SRE taxa and vice versa, it will be noted here.

C. Morphology Indicators: • A suite of morphological characters are characteristic of SRE taxa. • Where morphological characters are known to be associated with SRE taxa and vice-versa, it will be noted here.

D. Molecular Evidence: • If molecular work has been done on this taxon (or a close relative), it may reveal patterns congruent or incongruent with SRE status.

E. Research & Expertise: • Previous research and/ or WAM expertise elucidates taxon SRE status. • This category takes into account the expert knowledge held within the WAM.

Warramboo Troglofauna Assessment

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Appendix 6

Desktop Review: Previous Studies and Comparison

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx

Warramboo Troglofauna Assessment

Study Current (2006) Biota Bennelongia (2010) (2012a) Biota (2014) MWH (2014b) Biota Phase 1 2 1 2 3 4 5 6 1 1 1 1 Area Surveyed Warramboo Warramboo Mesa Mesa A, B, Mesa A, Mesa Mesa Mesa Dinner Mesa A Mesa A Tod Bore, A, G, C, F, G, K, Warramboo A A A Camp, and B and B Hubert H, J 2402e, Todd Congo Well, Bore, Bore Congo Warramboo Bore, Highway Deposit Survey Timing 3 Jun' - 8 Aug' 8 Aug' - 2 21 7 Apr' - 28 26 Jul' - 8 3 Jun' 19 11 Apr' 22 Sep' – 23 May - 25 Jun' 16 Oct' - 8 2015 Oct' 2015 Nov' Jul' 2005 Sep' 2005 - 3 Dec' - 23 18 Nov' 20 Jul' - 5 Sep' Dec' 2013 2004 Aug' 2006 - May 2010 2012 2014 - 10 2006 6 Feb' 2007 Jan' 2007 2005 Rain data (mm) Rain during Sampling 14.2 0 99.4 44 0 0 10.2 14.4 0 51.6 0 7.6 Rain 3 months preceding 254.8 28.4 1.8 627.4 48.4 191.4 23.8 76.2 26.8 47.4 110 1 Sites Sampled Trapped 33 33 25 88 73 31 31 36 40 46 51 116 Number of traps 69 57 85 291 221 89 84 109 80 135 158 301 Scraped 8 0 0 0 0 0 0 0 40 0 50 65 Sites Overlapping current study area Trapped (# traps) 33 33 3 (8) 4 (11) 19 (55) 0 17 (44) 17 (45) 40 3 (11) 4 (11) 116 Scraped 8 0 0 0 0 0 0 0 40 4 65 Collections Araneae X X X X Blattodea X X X X Coleoptera X X X X X Diplura X X X X X X X X X X Geophilamorpha X Isopoda X X X X X X

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx Warramboo Troglofauna Assessment

Study Current (2006) Biota Bennelongia (2010) (2012a) Biota (2014) MWH (2014b) Biota Phase 1 2 1 2 3 4 5 6 1 1 1 1 Palpigradida X Pauropoda X Polydesmida X X X X X X X X Pseudoscorpiones X X X X X X X X X X X Schizomida X X X X X X X X X X Scolopendromorpha X X X X X X X Symphyla X X Thysanura X X X X X X X X X X

Cube:Current:1080 (Robe Valley Subterranean Fauna 2015):Documents:Warramboo Troglofauna:Troglofauna Rev A.docx