SCIENCE AND CONSERVATION DIVISION

Aquatic Fauna Survey at Mandora Marsh (Walyarta) in September 2015

Prepared for Parks and Wildlife’s Kimberley Region

Kirsty Quinlan, Adrian Pinder and Loretta Lewis Wetlands Conservation Program Department of Parks and Wildlife

November 2016

1

Acknowledgements

We respectfully acknowledge the Nyangumarta as the traditional owners of the land on which this survey was conducted. We are grateful to you for imparting your cultural knowledge, for introducing us to the springs and wetlands of Walyarta and for assistance during the survey.

The following are also thanked for their assistance at various stages of this survey:-

Tracy Sonneman and staff from the Kimberley Region for co-ordinating the survey and providing logistical support.

Russell Shiel (University of Adelaide) – specialist identification of micro-fauna (including Cladocera, Rotifera and Protista)

Stuart Halse & Jane McRae (Bennelongia Environmental Consultants) – specialist identifications of Ostracoda and Harpacticoida

Tom Weir (CSIRO National Research Collections Australia, Canberra) – specialist identification of several Hemiptera

Chris Watts (South Australian Museum) – specialist identification of several Coleoptera

The project was proudly supported by BHP Billiton as part of Eighty Mile Beach and Walyarta Conservation Program (via the Department’s West Kimberley District Office).

Recommended Reference: The recommended reference for this publication is: Quinlan K, Pinder A, Lewis L (2016). Aquatic Fauna Survey at Mandora Marsh (Walyarta) in September 2015: Prepared for Parks and Wildlife’s Kimberley Region. Department of Parks and Wildlife, Kensington, Western Australia.

Title page photo: Montage of photos from Walyarta survey trip September 2015. Grants Spring and Salt Creek spring are featured.

2 Contents

1. Introduction ...... 6 1.1. Survey background ...... 6 1.2. Previous studies ...... 6 1.3. Ramsar listing and importance ...... 6 2. Overview of Mandora Marsh (Walyarta)...... 8 2.1. Location ...... 8 2.2. Land tenure ...... 8 2.3. Climate ...... 8 2.4. Wetland types ...... 9 3. Methods ...... 10 3.1. Site selection ...... 10 3.2. Water chemistry and habitat parameters ...... 12 3.3. Aquatic invertebrate sampling ...... 12 3.4. Invertebrate sample processing ...... 13 3.5. Data management ...... 13 3.6. Data analysis ...... 14 4. Results and discussion ...... 17 4.1. Water chemistry and habitat parameters ...... 17 4.2. Aquatic invertebrates ...... 21 4.2.1. Invertebrate diversity ...... 21 4.2.2. Community composition ...... 24 4.2.3. Aquatic invertebrates and environmental variables ...... 26 4.2.4. Biogeographic affinities of the 2015 fauna ...... 27 4.2.5. Groundwater species ...... 27 4.2.6. Noteworthy species ...... 30 5. Comparison between the present survey and the 1999 survey...... 33 References ...... 40 Appendix 1 – Water quality parameters and habitat information for springs and wetlands surveyed at Walyarta in 2015...... 44 Appendix 2 – Aquatic Invertebrate Species List (2015) ...... 46

3 Summary An aquatic survey of 15 springs and wetlands of the Mandora Marsh (Walyarta) system was undertaken in September 2015. The aim was to sample wetlands for water quality and aquatic invertebrates and to compare biodiversity at these springs with that documented during a survey of these springs undertaken in 1999 (Storey et al. 2011). Water quality parameters were comprehensively sampled at eleven springs, with seven of these springs differing very little in terms of the variables measured. At two of the springs (Saunders and Grants) higher levels of turbidity and nutrient (nitrogen and phosphorus) concentrations were recorded, indicative of enrichment by cattle. The two Salt Creek sites (18 and 19) were saline whereas the springs were mostly fresh to subsaline. Aquatic invertebrates were sampled at the same eleven springs, with at least 175 aquatic invertebrates collected in 2015, bringing the total number of species known from the springs to at least 216. The average species richness per spring was 46 and ranged from 11 (Stromatolite Pool) to 87 species (Eil Eil Spring). This richness is comparable to other similar surveys of springs in Western Australia. Communities were dominated by insect species (54.8%), followed by crustaceans (22.3%) and rotifers (10.2%). Most springs had similar invertebrate faunas, with Eil Eil and Little Eil Eil springs showing some differentiation from the rest. Invertebrate communities in the two Salt Creek sites had distinct communities reflecting their high salinity. The Walyarta aquatic invertebrate fauna collected in 2015 appears to be composed mostly of very widespread species (occurring across at least northern Australia or more widely). While only a few species are believed to be restricted to Western Australia, at least seven are currently known only from Walyarta, of which five were first collected in 2015. This is a significant number of potentially local endemics. However, since the ranges of many species are not known these patterns are only indicative at present. About 40% of species collected in 2015 were also collected in springs sampled by Pinder et al. (2010) in the Pilbara.

Notable records from the 2015 survey included a new cypridid ostracod collected from Fern Spring and Melaleuca Spring, which is being described. A Haliplus water beetle collected from Eil Eil Spring could not be matched with any described species and may also be new. Other potentially new finds include a chydorid water flea (cf. Celsinotum n.sp.) from Eil Eil and Little Eil Eil Springs, a Schizopera copepod from two springs and an Arrenurus water mite from four sites. The occurrence of the hemipteran Microvelia (Pacificovelia) lilliput was a significant record, representing a southern range extension for this species in WA (T Weir, pers. comm., 2016). These are in addition to two other locally endemic species, a syncarid crustacean and an assimineid snail, collected in 1999 (the latter recollected in 2015). A comparison with 1999 data revealed that species richness (at both the site and whole spring complex levels) was somewhat higher than in 1999 and that invertebrate composition was substantially different. A total of 134 aquatic invertebrate taxa were recorded from 10 sites in 1999, compared to a total of 175 taxa collected from 11 sites in 2015. Despite the differences between years, patterns of beta diversity across the springs showed some parallels. We do not know the temporal dynamics of the drying and wetting regimes of these wetlands, but there is evidence to suggest that these pools associated with the springs are temporary and highly 4 dynamic in nature. Together, the datasets from 1999 and 2015 present a more complete picture of the aquatic invertebrate diversity that is supported by these springs over a range of climatic and hydrological conditions.

5 Introduction

1.1. Survey background From the 31st August to 10th September 2015, the Department of Parks and Wildlife’s Kimberley Region led a multi-disciplinary team of ecologists and hydrologists to undertake a biodiversity survey and hydrological study of the Mandora Marsh (also known as Walyarta) area in northern Western Australia. Together with the Nyangumarta traditional owners, the team comprised individuals with expertise in terrestrial flora and fauna survey, aquatic fauna survey and hydrology. One component of this survey was an aquatic survey of the springs and wetlands of the Mandora Marsh (Walyarta) system. The aim of the aquatic survey was to sample a number of the mound springs and wetlands for water quality and aquatic invertebrates, and to compare biodiversity at these springs with that documented during a survey of these springs in 1999 (Storey et al. 2011). This report presents the aquatic sampling component of the survey and makes comparisons with the 1999 survey.

1.2. Previous studies The Walyarta area was first comprehensively surveyed for aquatic fauna in 1999 by Storey et al. (2011), as part of a larger biodiversity survey to document the terrestrial flora, vertebrate fauna and aquatic invertebrates of the area (Graham 1999; Start et al. 2008). Prior to this, only small collections of crustaceans and molluscs from several Mandora springs and Salt Creek had been made by Ponder (unpubl. data) in 1987. Between 1999 and 2000, waterbird surveys were conducted by air and from the ground at the marsh (Halse et al. 2005). More recently, in May 2008, one of the mound springs (Saunders Spring) was surveyed for aquatic fauna as part of the Inland Aquatic Integrity Resource Condition Monitoring (IAI RCM) project by the Department of Parks and Wildlife (Daniel et al. 2009).

1.3. Ramsar listing and importance The Mandora Salt Marsh, together with Eighty Mile Beach, was listed under the Ramsar Convention as a Wetland of International Importance in June 1990. The site itself comprises two separate areas, the beach (220 km) and associated intertidal mudflats from Cape Missiessy to Cape Keraudren, and the Mandora Salt Marsh, lying 40km inland (Hale & Butcher 2009) (Figure 1).

As well as being listed as a Ramsar wetland, the Mandora Salt Marsh is also listed on the Directory of important Wetlands in Australia (Environment Australia 2016), and included in one of Birdlife International’s ‘Important Bird and Biodiversity Areas’ (IBAs). The IBA includes a 3337 km2 area of Mandora Marsh and Anna Plains (BirdLife International 2016). Additionally, the Mandora Marsh springs are listed as Threatened Ecological Communities (TECs) in Western Australia.

6

Figure 1: Map showing the location of the Ramsar listed sites at Eighty-mile Beach (inclusive of Mandora Marsh) and Roebuck bay. Bioregions (IBRA7 http://www.environment.gov.au/land/nrs/science/ibra#ibra) are also illustrated.

7 2. Overview of Mandora Marsh (Walyarta)

2.1. Location The Walyarta system lies approximately 250 km south of Broome adjacent to Eighty-Mile Beach. The western edge of the marsh lies ~30 km inland from eighty-mile beach on the western edge of the Great Sandy Desert biogeographic region and sits between the local government Shires of Broome and East Pilbara at the transition between the Kimberley and Pilbara geographic Regions (Fig. 1).

2.2. Land tenure The majority of the land on which the Walyarta springs and wetlands occur had, until July 2015, been situated on the Anna Plains pastoral lease. Agreements have now been reached between the station owners and State Government of Western Australia, for a portion of this land (including the marsh area) to be excised from the pastoral lease and managed for conservation and cultural purposes. The remainder of the marsh area was already on Unallocated Crown Land.

2.3. Climate The survey area experiences a semi-arid monsoonal climate. It receives a median and mean annual rainfall of 338.3 and 376 mm respectively (data from Mandora Meteorological Station; Bureau of Meteorology 2016), falling predominantly in the summer wet season from December to March. Rainfall is highly variable between years (Fig. 2), owing largely to frequent tropical cyclones that cross the northern WA coastline in the vicinity of eighty-mile beach. This cyclonic activity has historically resulted in extensive flooding of the Walyarta and surrounds (1942, 1980, 1997, 1999, 2000 and 2011), with annual rainfall in three of these years above 800 mm.

When first surveyed in 1999, Walyarta and surrounds were extensively inundated and supported very large numbers (~490,000) of waterbirds (Halse et al. 2005). In the 5 years prior to the survey (1994 – 1999), a tropical cyclone had crossed the coast each year and annual rainfall for each of those years was well above the mean (Storey et al. 2011; Fig. 2). In the year preceding the 1999 survey (Oct 1998 – Oct 1999) the area had received 982 mm of rainfall (Bureau of Meteorology 2016). By contrast, the 5 years prior to the September 2015 survey were not as wet. Some flooding was documented in 2011 (Johnstone et al. 2013) and rainfall was also high (750 mm) in 2013 but total rainfall in 2014 and 2015 was below the mean of 376 mm (Bureau of Meteorology 2016; Fig. 2). In the year preceding the 2015 survey (Aug 2014 – Aug 2015) the area had received only 327 mm of rain (Bureau of Meteorology 2016; Fig. 2).

8

Figure 2: Total annual rainfall from Mandora meteorological station for the period 1914-2015. Rainfall for the 5 years preceding both the 1999 and 2015 surveys are shown in pink. Information from the Bureau of Meteorology, accessed August 2016.

2.4. Wetland types Walyarta and surrounds covers an area approximately 105 000 ha in size and comprises several wetland types. There are two large seasonal wetlands which are usually filled following cyclonic rainfall events. Lake Walyarta (approximately 8000 ha) is on the western edge of the survey area and extends eastwards for approximately 30km (Graham 1999). The eastern wetland is more of a braided drainage line with patches of vegetation and salt/clay pans. A saline creek (Salt Creek), approximately 10 km long, runs between the two lakes and is lined with the most inland occurrence of mangroves (Avicenna marina) in Western Australia (Graham 1999; Semeniuk & Semeniuk 2000; Storey et al. 2011). There is also a series of small permanent freshwater mound springs and salt springs scattered throughout the marsh system, varying in size from 0.1 ha to more than 11 ha (English et al. 2016). These mound springs typically have a raised central mound of peat which is frequently saturated and often has small pools of standing water (Hale & Butcher 2009). The mounds are generally surrounded by a partial or complete moat which is intermittently inundated. The depth of water at each mound spring varied from damp soil with little/no surface water present,

9 through to surrounding pools up to 40 cm deep (e.g. Grants Spring) (Hale & Butcher 2009; English et al. 2016). These springs typically support Sesbania formosa (dragon flower tree) and Melaleuca, often over Acrostichum speciosum fern or sedges (e.g. Fimbristylis or Typha) (English et al. 2016).

3. Methods

3.1. Site selection Site selection for the survey undertaken in September 2015 was largely based on the sites sampled in 1999 by Storey et al. (2011) to enable comparison of aquatic invertebrate communities between these two years. Aquatic invertebrates were sampled at 11 sites during the most recent survey (Figures 3, 4 & 5). This list included three additional sites which were not sampled for invertebrates in 1999. These sites were Grants Spring, Stromatolite Pool (Salt Creek) and the main Stockyard Spring. Several of the sites sampled in 1999 were not sampled in 2015. Of these, the large saline wetland (Lake Walyarta) and Coolabah Claypan were dry (latter confirmed by remote sensing) and Linear Spring and Sporobolus Spring were inaccessible in the time available. We also did not sample any of the bores. The full list of 2015 survey sites and their geographic coordinates are provided in Table 1 below.

Table 1: Aquatic survey sites sampled in September 2015 at Walyarta. Also indicated are the sites surveyed for aquatic invertebrates in 1999 (Storey et al. 2011).

Site sampled Site No. Date Site Name Latitude Longitude in 1999 for S E invertebrates

Sites comprehensively sampled for water chemistry and aquatic fauna

3A and 3B 4/09/2015 Fern Spring inner pool and outer pools * 19° 45' 59.9" 121° 23' 34.1"  4 4/09/2015 Melaleuca Spring * 19° 46' 10.5" 121° 23' 21.2"  5 3/09/2015 Saunders Spring * 19° 46' 57.1" 121° 20' 18.2"  6 6/09/2015 Little Eil Eil Spring * 19° 47' 35.0" 121° 26' 24.6"  7A and 7B 5/09/2015 Eil Eil Spring outer pool and inner pools * 19° 47' 48.2" 121° 26' 36.3"  9 2/09/2015 Top Spring 19° 48' 48.5" 121° 36' 50.8"  12 7/09/2015 Stockyard main spring 19° 45' 30.5" 121° 23' 12.1" - 17 5/09/2015 Bretts Spring 19° 47' 33.8" 121° 26' 57.3"  18 7/09/2015 Salt Creek Claypan Spring 19° 43' 56.1" 121° 28' 54.9"  19 1/09/2015 Salt Creek Stromatolite pool 19° 44' 35.4" 121° 32' 28.0" - 23 6/09/2015 Grants Spring * 19° 47' 00.7" 121° 21' 22.0" -

Basic water chemistry only was measured

13 7/09/2015 Small spring with mangrove at stockyard 19° 45' 30.5" 121° 23' 15.6" - 14 7/09/2015 Mangrove/Typha spring 100m W of stockyard 19° 45' 31.2" 121° 23' 08.2" - 15 7/09/2015 Small spring 200m ESE of stockyard 19° 45' 34.1" 121° 23' 15.2" - 16 7/09/2015 Spring with Typha and moat 150m W of stockyard 19° 45' 30.0" 121° 23' 05.3" - *Denotes sites where microfauna was also extracted from a one meter deep core sunk into the peat

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Figure 3: Location of 15 aquatic sites sampled at Walyarta in September 2015. For corresponding site names, refer to Table 1.

11 3.2. Water chemistry and habitat parameters At each of the 15 sites sampled in 2015, basic water chemistry measurements (turbidity (NTU), electrical conductivity (µS cm-1), salinity (g L-1), water temperature (°C) and pH were measured in-situ using hand held calibrated meters (TPS WP-81 and WP-88). The probes of the meters were suspended in the water column approximately mid-way between the water surface and the sediment.

At 11 of these sites (listed in Table 1), more comprehensive sampling for water chemistry and aquatic fauna was undertaken. A water sample (~750 ml) was collected approximately 15 cm below the water surface (or slightly less for some of the shallower springs) for use in more detailed water chemistry analyses by the Chemistry Centre of Western Australia. Samples being analysed for total filterable nitrogen and total filterable phosphorus were first passed through a 0.45 µm pore size filter in the field and then frozen. Chlorophyll and phaeophytin were also measured at the Chemistry Centre using phytoplankton that had been retained on glass-fibre filter paper through a water filtration tower.

Habitat and substrate information was recorded at each of these eleven comprehensively sampled sites, including an estimate of the percentage cover of emergent and submerged aquatic plants in the area sampled for invertebrates. Sediment was collected in a 1L pot and particle size analysis was undertaken at the Chemistry Centre.

3.3. Aquatic invertebrate sampling Two aquatic invertebrate sweeps were taken at each of the 11 main survey sites (both sweeps covering the same area of the wetland). A plankton sample, using a 50 µm mesh net to sample invertebrates in the water column, and a benthic sample, using a 250 µm mesh net to sample larger animals in the water column and benthic habitats within wadeable depth (e.g. open water, submerged vegetation, sticks/logs/leaf litter). These two sweeps ensured that both micro- and macro-invertebrates were captured. Due to the small size of some of the springs and the limited volume of surface water available to sample, a sweep length less than the standard 50 m was employed. Site number and sweep length for the 2015 survey is recorded below (Table 2). A maximum water depth was recorded in the sampling area where the invertebrate sweeps were collected. Coarse inorganic sediment and coarse organic matter were removed prior to sample preservation by washing debris and elutriating in buckets before passing the water back through the net. Samples were then preserved in 100% ethanol in the field and taken back to the laboratory for processing.

Interstitial fauna was sampled at six of the mound springs (Table 1) following methods outlined in Storey et al. (2011). At each of these sites, a split corer was used to extract a one metre deep core and the resultant hole was allowed to fill with water. The water was then pumped out through a 110 µm mesh net and the contents placed into a 2L pot and preserved with 100%

12 ethanol. This sample was taken back to the laboratory for processing and identification of microfauna.

Table 2: Sweep lengths for aquatic fauna sampling at Walyarta (September 2015).

Site 3A 3B 4 5 6 7A 7B 9 12 17 18 19 23 Plankton sweep length (m) 5 30 10 25 30 30 50 50 25 30 50 30 30 Benthic sweep length (m) 7 30 20 20 30 50 50 50 25 30 50 30 30

3.4. Invertebrate sample processing Plankton samples were washed with tap water and sieved through 250 μm, 90 μm and 50 μm sieve sizes. Only the 250 μm, and 90 μm fractions were examined further under the dissecting microscope, with representatives of each discernible species (mainly macrofauna) picked out and preserved in 100% ethanol. The majority of the microfauna including the Protozoa, Rotifera and Cladocera were left in the bulk sample, which was then sent to a consultant in Adelaide for specialist identification of these groups.

The benthic samples were washed with tap water and sieved through 2 mm, 500 μm and 250 μm sieve sizes. Each sieve fraction was examined and representatives of each discernible species were picked out under a dissecting microscope and preserved in 100% ethanol. All taxa were identified to the lowest taxonomic level possible using keys and voucher specimens and undescribed taxa were assigned morphospecies names based on previous survey work by Parks and Wildlife (Wetlands Conservation Program). A select group of ostracods from the survey and all of the harpacticoid copepods were sent to a consultant for specialist identification. Several specimens from the orders Hemiptera and Coleoptera were also sent to relevant experts (Tom Weir and Chris Watts) for identification confirmation. For a select group of beetles (Berosus), DNA barcoding (a technique where a short DNA sequence from an unidentified specimen is compared to a library of known DNA sequences) was used to improve the taxonomic resolution of this group to species level at a number of wetlands.

A survey specific reference collection was prepared and will be lodged with the Western Australian Museum.

3.5. Data management

Prior to data analysis, 2015 species presence/absence data was edited to remove most identifications above family level, and specimens that were unable to be identified morphologically due to gender or life-stage constraints. When comparing the 1999 and 2015 aquatic invertebrate dataset, some of the data needed to be grouped to genus or family level because of differences in taxonomic resolution between years. This occurred for some Diptera groups such as Ceratopogonidae, Ephydridae and some Chironomidae.

13 Aquatic invertebrate data from interstitial samples was excluded from both the 2015, and the combined 1999/2015 datasets for analyses. However, this information contributed to the final species list.

3.6. Data analysis

Multivariate analyses were undertaken using the PRIMER (v7) software package (Clarke & Gorley 2015). Patterns between sites, based on environmental variables (water quality and habitat-sediment data) were explored using cluster and ordination techniques. Firstly, all variables were viewed in the ‘Draftsman Plot’ function in PRIMER. For pairs of variables that showed a strong correlation (>0.98), one of each pair was excluded from the analyses. Data was transformed (to make distributions more normal) where required and then range standardised. Hierarchical agglomerative clustering (UPGMA) was used to group sites with similar environmental parameters, and a similarity profile permutation test (SIMPROF) was used to test the statistical support of the branches produced in the classification (Clarke et al. 2008). A non- metric multidimensional scaling (nMDS) ordination (Clarke & Gorley 2015) was then used to visualise these groups in a two-dimensional plot. The cluster analysis and ordination for this dataset were based on Euclidean distance.

For aquatic invertebrate presence/absence data, a cluster analysis was first undertaken, based on Bray-Curtis similarity matrices (Bray & Curtis 1957). The removal of singleton taxa (species which only occurred at one site) made no difference to the classification and so these were left in. The SIMPROF routine was used to test the statistical support of branches produced in the classification. An nMDS ordination was used to visualise the similarity of invertebrate community composition between sites in a two-dimensional plot. The similarity percentage (SIMPER) routine (limited to 70% cumulative similarity) was then used to identify those species contributing most to the similarity/dissimilarity between sites. The relationship between invertebrate data and environmental parameters was assessed using the BEST routine (Clarke et al. 2008). The BIO-ENV function was used to calculate the smallest sub-set of environmental variables that explained the greatest percentage of variation in the ordination patterns.

To explore differences in aquatic fauna assemblages between survey years (1999 and 2015), additional analyses were undertaken. An nMDS ordination was used to visualise the similarity of invertebrate community composition between sites and years, in a two-dimensional plot. For a priori groups (e.g. year), analysis of similarity (ANOSIM) was conducted to determine if the groups were significantly different from one another.

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Figure 4: Walyarta aquatic survey sites in 2015 (sites 3A – 7A). A: Fern Spring inner pool (3A), B: Fern Spring outer pools (3B), C: Melaleuca Spring (4), D: Saunders Spring (5), E: Little Eil Eil Spring (6), F: Eil Eil Spring outer pool (7A).

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Figure 5: Walyarta aquatic survey sites, continued (sites 7B – 23). A: Eil Eil Spring inner pools (7B), B: Main Stockyard Spring (12), C: Bretts Spring (17), D: Salt Creek Spring (18), E: Stromatolite Pool (19), F: Grants Spring (23).

16 4. Results and discussion

4.1. Water chemistry and habitat parameters Water chemistry (basic and comprehensive) and habitat data for each of the survey sites is provided in Appendix 1. Cluster analysis and ordination of the 11 sites at which comprehensive water quality and habitat information was collected, identified three SIMPROF supported cluster groups based on environmental variables (Fig. 6). The largest group comprised most of the mound springs, with the exception of Saunders Spring (5) and Grants Spring (23), which grouped separately. Both of the salt creek sites (18 and 19) also formed a separate grouping.

2D Stress: 0.1 Group 19 a b c

4 23

17 7A 5 18

12 3A

3B 6

9

Figure 6: Two-dimensional non-metric multidimensional scaling (nMDS) ordination of 11 sites comprehensively sampled for water chemistry and habitat data in September 2015. Ordination is based on selected, transformed, and normalised water quality and habitat (sediment) data.

Salinity was variable, highest for the Salt Creek sites (18 and 19; Fig. 7), with values of 35.7 g.L-1 and 37.1 g.L-1 respectively. Four of the springs (3, 4, 6 and 17) were sub-saline (3 – 10 g.L-1) and five were considered fresh (<3 g.L-1). Ionic composition showed most sites to be dominated by Na and Cl ions (Appendix 1). The majority of the springs and pools were clear, with field turbidity < 10 NTU. The exceptions were Saunders Spring (5) and Grants Spring (23) with higher turbidity values of 49 and 170 NTU respectively (Fig. 7), which may be contributing to their separation from other springs in Figure 6. Both of the Salt Creek sites (18 and 19) had very low colour (<10

17 TCU), but all of the mound springs were more coloured with TCU values ranging from 40 – 280 TCU, likely reflecting more vegetation debris and particulate organic matter in the springs. The highest colour value (450 TCU) was recorded at Grant Spring (site 23) (Appendix 1). Most sites were alkaline (pH 7.4 - 8.99; Fig. 7) which is reflective of the soils and underlying groundwater in the area. The exception was one of the small springs surrounding the main stockyard (site 16) which had a pH of 6.63 (Appendix 1).

Nutrient concentrations at the mound springs were variable. Total nitrogen (TN) concentration ranged from 0.35 - 4.6 mg.L-1 (mean 1.5 ± 1.2 SD). The two highest TN concentrations were at Saunders Spring (2.7 mg.L-1) and Grants Spring (4.6 mg.L-1). Total Filterable Nitrogen (TFN) ranged from 0.34 - 2.3 mg.L-1 (mean 0.99 ± 0.6 SD) with the highest concentrations recorded again at sites 5 and 23 (Fig. 7). Total Phosphorus (TP) concentrations ranged from 0.005 - 0.73 mg.L-1 (mean 0.2 ± 0.2 SD). The two highest TP concentrations occurred at Saunders Spring (0.73 mg L-1) and Grants Spring (0.54 mg L-1). Total Filterable Phosphorus (TFP) ranged from 0.005 mg.L-1 to its highest concentration of 0.4 mg.L-1 (mean 0.07 ± 0.11 SD) at Saunders Spring (Appendix 1; Fig. 7). These high nutrient concentrations at sites 5 and 23 may be another factor contributing to the separation of these sites in Figure 6.

Surveys of similar springs, such as the organic mound spring complex near the town of Three Springs (200 km NE of Perth), also revealed high nutrient levels at some sites (Pinder & Leung 2010). Total nitrogen concentrations across six of those mound springs ranged from 0.07 - 6.3 mg.L-1 (mean 1.6 mg.L-1), and total phosphorus from 0.005 - 0.3 mg.L-1 (mean 0.2 mg.L-1). These springs are also surrounded by agricultural land which could account for these higher concentrations. Additionally, a survey of five selected Pilbara springs also showed variation in nutrient concentrations, with an average total nitrogen concentration of 1.9 mg.L-1 and an average total phosphorus concentration of 0.01 mg.L-1 (Halse et al. 2002).

Most sites had low phytoplankton concentrations (chlorophyll-a < 5 µg.L-1). Three sites including Melaleuca Spring, Grants Spring and Saunders Spring, did have some obvious algal turbidity and marginally higher chlorophyll-a values of 10, 8 and 7 µg L-1 respectively (Fig. 7; Appendix 1).

Australian water quality guidelines (ANZECC & ARMCANZ 2000)for arid-zone wetlands suggests trigger TN concentrations of under 1.2 mg.L-1, trigger TP concentrations of under 0.05 mg.L-1 and trigger chl-a concentrations of < 0.01 mg.L-1. It is important to note that the ANZECC/ARMCANZ guidelines were based on minimal background data (little data for north-western Australia was available at the time) and are being revised. Nonetheless, our results suggest some nutrient enrichment of the Walyarta springs, particularly Saunders Spring (5) and Grants Spring (23), probably associated with the presence of cattle (which were observed at both springs). The higher turbidity at sites 5 and 23 probably also reflect phytoplankton growth and physical disturbance of sediments by cattle.

18

Salinity (g L -1)

Turbidity (NTU)

pH

-1 Total Filt N (ug L )

Total Filt P (ug L -1)

Chlor-a (ug L -1)

Figure 7: Bar chart plots of selected water quality parameters recorded for 11 sites sampled at Walyarta in 2015. The 3 main groupings from the nMDS ordination of water quality parameters (Fig. 6) are also shown, delineated by the red lines in the plot.

19 Water depth at the mound springs was low: <40 cm. Estimated submerged macrophyte cover was highest at Eil Eil Spring pools (7B) with approximately 90% cover (Appendix 1) and lowest at Fern Spring inner pool (3B), where the substrate was almost bare. These sites also corresponded with highest and lowest invertebrate richness respectively. Emergent macrophyte cover was generally minimal (<5%) at most springs. However, at Grants Spring there were patches of Typha and at Bretts Spring patches of Fimbristylis, which each covered approximately 25% of the area sampled for aquatic fauna (Fig. 5; Appendix 1). The importance of macrophytes in providing habitat, protection and a food source for invertebrates has been documented previously (Boulton & Lloyd 1991; Sheldon et al. 2002), as has the relationships between invertebrate assemblage richness and macrophyte abundance (Wollheim & Lovvorn 1996; Pinder et al. 2010). However, the relationship between macrophyte cover and species richness in this survey was not linear, perhaps because there are a number of variables contributing to species richness at these wetlands.

Sediment particle size composition at each of the 11 sampled sites is illustrated in Figure 8. Sand was the dominant size fraction at almost all sites (mean contribution of 53%), followed by silt (mean 29%) and then clay (mean 18%). Saunders Spring (site 5) was an exception with marginally higher silt content (40%) than sand (37.5%), as were Fern Spring inner pools (site 3A) and Salt Creek Spring (site 18) which both had very low percentages of silt (4% and 6% respectively). Both of these latter sites also had the highest sand content of all sites, with 84.5% and 85.5% respectively. Stones were present at each site, however the percentage contribution of these was minimal, ranging from 0.05% - 3.1% (Appendix 1).

Figure 8: Sediment particle size composition for 11 aquatic sampling sites (including subsites) at Walyarta in 2015.

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4.2. Aquatic invertebrates

4.2.1. Invertebrate diversity

At least 175 species of aquatic invertebrates were collected from 11 wetlands in September 2015 (Appendix 2). The average richness was 46 taxa ± 6.6 (SE) per wetland (excluding interstitial samples). The highest richness was at Eil Eil Spring (site 7), with 87 taxa, and the lowest at Stromatolite Pool (site 19) with 11 taxa (Fig. 9). Diptera were the most diverse group, with 34 taxa, 12 of which were chironomids. The number of Diptera taxa listed is likely to be an underestimate of the total number of species present in these wetlands, as several of the families and genera (i.e. Ephydridae spp., Anopheles spp., Tanytarsus spp. and Monohelea spp.) likely contain multiple species. At present, there are taxonomic limitations to identifying some of these groups to species level, and in these cases molecular methods could be employed to better resolve these problem groups.

Figure 9: Species richness at wetlands sampled in 2015. Individual totals for subsites at sites 3 and 7 are also shown (light grey).

Almost half of these taxa (41.5%) were only recorded in one of the 11 sites, with ~15% present in more than half (6 or more) of the sites. The presence and abundance of protists, rotifers and cladocera in these samples was low, with protists and rotifers being completely absent from four or more sites (e.g. 3, 4, 9 and 17), and cladocera absent from six (4, 5, 12, 18, 19 and 23). Small springs do tend to have a less diverse microinvertebrate fauna compared to larger open lentic wetlands.

21 Species richness at Walyarta can be compared with other similar surveys of springs within the State. At Three Springs, 200 km north-northeast of Perth, at least 124 species have been recorded from a small group of five organic mound springs to date, with an average richness of 29 species per spring (Pinder & Leung, 2010). Forming a subset of a much wider survey of the Hutt catchments (Pinder et al. 2012) was a group of freshwater springs, spring-fed creeks and seepages. This subset contained 151 species, with an average richness of 47 species per spring (Pinder et al. 2012). Species richness per spring in all three of these surveys was much lower than recorded for springs in the Pilbara. A survey of five springs by Halse et al. (2002), documented 159 species with an average of 60.4 species per spring, and a more extensive survey of 16 Pilbara springs by Pinder et al. (2010) recorded nearly 600 species. The latter were collected over two seasons with 80% occurring in both seasons (465 in autumn and 494 species in spring). In these springs the average species richness per spring was 104 species in autumn and 114 in spring. Pilbara springs were associated with more permanent creek systems and larger drainage networks allowing greater invertebrate colonisation potential, whereas the Mandora, Hutt and Three Springs springs were more isolated swampy wetlands.

Interstitial samples were collected from six of the mounds springs during this survey. These were Fern, Melaleuca, Saunders, Little Eil Eil, Eil Eil and Grants springs. Species richness was highest at Eil Eil Spring with 14 species and lowest at Melaleuca Spring with only 2 species. A total of 31 species were collected from interstitial samples and was composed of the following groups: Copepoda (7 species), Diptera (6), Ostracoda (5), Oligochaeta (3), Rotifera (3), Cladocera (2), Acarina (2), Gastropoda (2), Nematoda and Turbellaria. Some of these are groundwater species discussed below.

A selection of aquatic fauna collected during this survey is shown in Figure 10. The fauna includes some noteworthy species which are discussed in more detail below in Section 4.2.6.

22

A B

C

D

E

Figure 10: Aquatic invertebrates collected in 2015. A. Gyraulus sp. (site 9); B. Oecetis sp. (site 4); C. cf. Celsinotum n.sp (site 6) (image RJ Shiel); D. Anopheles sp. (site 3B); and E. Triplectides sp. in its case and an attached pupal Ephydridae in its own case (site 23).

23 4.2.2. Community composition Insects accounted for 54.8%, crustaceans (22.3%), rotifers (10.2%), and remaining groups, 12.7% of the total fauna collected from the 11 sites in 2015 (Fig. 11).

Figure 11: Pie chart showing the percentage composition of aquatic invertebrate groups collected from 11 wetlands at Walyarta in 2015. This chart excludes Protista.

An nMDS ordination showing all 2015 survey sites, based on species presence/absence data is shown in Figure 12. This analysis attempts to place each site on the plot so that the distances between sites (and subsites), best represents the overall similarity/dissimilarity of their invertebrate communities. Four groupings (groups A, B, C and D) are identified in the ordination based on significant cluster analysis groupings.

24

Figure 12: Two-dimensional non-metric multidimensional scaling (nMDS) ordination of Walyarta wetland sites based on species presence/absence data collected in 2015. The ordination shows four groupings.

Group A: This ‘group’ is composed of only one site, Fern Spring inner pool (3A). This small pool was situated deep on the densely vegetated Fern Spring peat mound (Fig. 4A). Site 3A is one of two subsites at Fern Spring, but it is clear from the ordination that this small pool was quite different from the suite of pools lying around the outer edge of the same spring (site 3B). When invertebrate data from these subsites were combined, Fern Spring (site 3A+B) grouped with sites from ‘group B’ in the ordination. The dissimilarity observed between 3A and other sites could be due to the low species richness of this site, itself reflecting low habitat diversity and that only a small sample was collected due to the small size of the pool (<2 m in diameter) (Table 2; Fig. 9). Sites 3A and 3B had similar water quality but 3A had sandier sediments (Fig. 6 and 8). Group B: This ‘group’ consists of Fern Spring outer pools, Melaleuca Spring, Saunders Spring, Eil Eil Spring outer pool, Top Spring, Main Stockyard Spring, Bretts Spring and Grants Spring (3B, 4, 5, 7A, 9, 12, 17 and 23). This was the largest group comprising eight samples from eight different springs, with a total of 128 species (excluding interstitial samples). These sites tended to be more open pools on the fringes of the mound springs (with the exception of Top Spring). Species strongly characteristic of this group included the copepod Mesocyclops brooksi, Anopheles mosquitoes, Monohelea ceratopogonids, the chironomid Procladius paludicola, damselfly Ischnura aurora aurora, dragonfly Orthetrum caledonicum and soldier fly larvae (Stratiomyidae).

25 When invertebrate data from subsite 7A is combined with 7B, it groups together with site 6 in ‘group C’ (similar to the position of 7B currently).

Group C: This ‘group’ includes Little Eil Eil Spring and Eil Eil Spring inner pools (6 and 7B). These two sites were both heavily shaded complexes of interconnected pools, unlike the more open pool sites of ‘group B’. Eighty-six species were recorded from these two sites, with 45 species (~52%) occurring in both sites. Eleven species contributed most to the discrimination between these two groups and included the protists Arcella discoides, Arcella sp. a, and Centropyxis aculeata, rotifers Lecane bulla and Lecane stenroosi, cladoceran cf. Celsinotum n.sp oligochaete Allonais ranauana, water mite Oribatida type 2, chironomid Pentaneurini sp., Scirtidae beetle larvae and the hemipteran Microvelia (Pacificovelia) lilliput.

Group D: This ‘group’ includes both of the Salt Creek sites (18 and 19) with 17 and 11 species respectively and 18 in total, with ten recorded from both sites. This low richness reflects the high salinity of these sites. Salt Creek Spring (18) and Stromatolite Pool (19) had the two highest salinity readings of all 15 sites measured for basic water chemistry parameters with 35.7 g.L-1 and 37.1 g.L-1 respectively (Fig. 7; Appendix 1). Six taxa (Lecane grandis, Lecane ungulata, Diacypris spinosa, Robertsonia propinqua, Helochares tatei and Tanytarsus barbitarsis) were restricted to group D. This small group of species, plus Apocyclops dengizicus from site 18 are well-known and widespread halophilic or halotolerant species (Pinder et al. 2012).

This analysis suggests that most of the springs have similar faunas, but that the Eil Eil spring complex (sites 6 and 7) may be somewhat different to the others (although the proximity of groups B and C suggests the difference is not great). Salt Creek clearly has a very different fauna reflecting its very different habitat.

4.2.3. Aquatic invertebrates and environmental variables

A BEST (BIO-ENV) analysis (PRIMER-E; Clarke & Gorley 2015) was used to investigate the relationship between aquatic invertebrate communities and environmental variables at 11 of the wetlands. The analysis suggests that a combination of three environmental variables (salinity, % K, and % silt) best explains the patterning of invertebrate community composition at Walyarta (Table 3). Indeed, Salt Creek Spring and Stromatolite Pool (sites 18 and 19) recorded high salinity values (35.7 g.L-1 and 37.1 g.L-1 respectively) in contrast to all of the remaining springs (range 1.23 – 7.4 g.L-1), and is likely driving the different aquatic invertebrate assemblages observed at sites 18 and 19 (Figure 12). The percentage of silt in the sediments at sites 18 and 3A were the lowest of all sites sampled, with both sites having a very high sand content. Most of the springs, however, have similar faunas and are reasonably similar in terms of water quality parameters (apart from slightly elevated turbidity and nutrients at sites 5 and 23), so apart from an obvious difference in salinity, very strong correlations with other environmental variables wouldn’t be expected.

26

Table 3: BEST (BIO-ENV) results from PRIMER showing which environmental variables (up to 5 variables) best explain the biotic patterns observed at Walyarta.

Variable No. of variables Correlation combinations

1 0.796 Salinity 2 0.789 Salinity, % K 3 0.826 Salinity, % K, % Silt 4 0.817 Salinity, % Ca, % K, % Silt 5 0.822 Salinity, pH, % Ca, % K, % Silt

4.2.4. Biogeographic affinities of the 2015 fauna Walyarta appears to represent a typical desert aquatic invertebrate fauna that favours habitat generalists and/or taxa with high vagility capable of moving between wetlands (Boulton et al. 2006; Storey et al. 2011). Most of the species have continental (or broader) distributions (Fig. 13), although we could not determine the broader distributions for just over a third of species due to taxonomic impediments, incomplete survey and data availability. Distributions have been determined from Parks and Wildlife data, published information and the Atlas of Living Australia. The 12% of species classed as Austro-pacific occur in Australia (especially northern Australia) but are also variously distributed through Papua New Guinea, Indonesia, the Pacific and New Zealand, but not more broadly in the Oriental region. This includes many of the dragonflies plus a few hemipterans and beetles and the mosquito Culex (Culex) annulirostris. Many of the 28% of species classed as ‘wider than Austro-pacific’ occur in the broader Oriental region but a few are even more widespread. Northern Australian species, representing 5% of the fauna, are all beetles or hemipterans, with the exception of the mosquito Culex starckeae. Twelve percent of species are widely distributed across Australia but not known from other regions.

Thirteen species (or at least their genera), representing about 8% of the fauna, are well enough known to suggest they are restricted to Western Australia. Of these, six are currently known only from Walyarta. These are the undescribed assimineid snail, a new genus and species of cyprinid ostracod that is being described, the Celsinotum water flea, a Schizopera copepod (sp. B23), an Arrenurus water mite and a Haliplus beetle (nr bistriatus). Remaining species that may be restricted to Western Australia are primarily copepods (especially groundwater associated Nitokra and Schizopera), but also an undescribed member of a complex of Heterocypris ostracods, the darwinulid ostracod Vestalenula matildae (a stygophilic species) and the water mite Arrenurus balladoniensis.

27

Figure 13: Biogeographic affinities of aquatic invertebrates from A) Walyarta springs and B) Pilbara springs sampled by Pinder et al. 2010.

Biogeographic affinities of the Walyarta fauna can be contrasted to that occurring in 16 springs sampled during the Pilbara Biological Survey (Fig. 13) by Pinder et al. (2010). Of the 175 species collected at Walyarta in 2015 about 40% were also collected in these Pilbara springs. The main difference in biogeographic affinities between Pilbara and Walyarta springs is that the Pilbara spring fauna had a much smaller proportion (27% compared to 40%) of species that also occur outside of Australia. The Pilbara springs had correspondingly greater proportions of species with northern or Western Australian (including north-western) ranges (11% each). Sixty percent of the species from Pilbara springs assumed to be Western Australian endemics were suspected of being restricted to the Pilbara by Pinder et al. (2010). Despite the proximity of Walyarta to the Pilbara only one such species was recorded from the Walyarta springs (the groundwater

28 associated ostracod Vestalenula matildae), although some of the Cypretta ostracods may be the same as some Pilbara species. The virtual absence of these presumed Pilbara endemics most likely reflects the different nature of the Walyarta and Pilbara springs. None of the Pilbara springs were mound springs. Instead, most were either discharges from rocky geology, usually forming semi-permanent creeks and pools, or were fed by permanent discharges within broader river beds. All were part of larger freshwater drainage networks providing greater colonisation potential and many had a wider variety of habitats than in the Walyarta springs, usually including some flowing water supporting lotic (flowing water) species. Many of the suspected Pilbara endemics were either groundwater species (which tend to have restricted distributions and limited dispersal potential) or tend to occur in riverine habitats rather than lentic wetlands. At least some of the Walyarta springs lack surface water at times which would also limit species richness by elimination of species not able to take refuge in the moist peats of the mounds.

4.2.5. Groundwater species

Several of the collected invertebrates are closely associated with groundwater. Schizopera is a primarily marine copepod genus that has diversified in subterranean waters in north-western and inland Western Australia (Karanovic 2006; Karanovic & Cooper 2012). Three species were collected in 2015 but none in 1999. One of these, Schizopera B23 from interstitial samples taken from Saunders and Little Eil Eil springs has not been collected elsewhere. Schizopera B24 was also collected only from interstitial samples (Fern Spring and Eil Spring) whereas Schizopera B22 was collected from the small pool on Fern Spring. The two Nikokra species (B04 and B05) are probably also stygophiles. These harpacticoid copepods belong to a ‘Nitokra lacustris species complex’ (J McRae, pers. comm., 2016) that is yet to be resolved. The Ameiridae sp. collected in 1999 could have been Nitokra. The cyclopoid Halicyclops spinifera is also closely associated with groundwater or groundwater discharge sites but has rarely been collected in Western Australia. Three other cyclopoids, Apocyclops dengizicus, Microcyclops varicans and Mesocyclops brooksi, are regularly collected in groundwater but are also widespread in surface water in Western Australia, as is Metacyclops cf. mortoni collected in 1999. Darwinulid ostracods are primarily stygophilic and the three species collected from Walyarta have been recorded in groundwater elsewhere in north-western Australia and two (Vestalenula marmonieri and Penthesilenula brasiliensis) are globally distributed. Vestalenula matildae, collected in an interstitial sample from Grants Spring is known only from north-western Australian groundwater (Halse et al. 2002; Pinder et al. 2010). Enchytraeid oligochaetes were primarily collected in the interstitial samples and the family regularly occurs in groundwater in WA. However, they could easily be soil dwelling species rather than stygophiles. Finally, the syncarid, Kimberleybathynella mandorana, collected in an interstitial sample from Fern Spring in 1999, is certainly a groundwater species. It is likely that further groundwater species would be found with further survey and that a larger stygofauna community would be inhabiting the associated aquifers.

29 4.2.6. Noteworthy species

Most of the aquatic invertebrate fauna collected in 2015 are common and widespread species. However, a number of noteworthy taxa were collected in 1999 and 2015 and these are discussed below. At least seven species are currently known only from Walyarta. Rotifera Lecane grandis, recorded from Salt Creek Spring (site 18) is a halophilic (salt-loving) species rarely recorded in Australia. While it has a global distribution (Segers 2007) there are few other Australian records: Kakadu (R Shiel, pers. comm., 2016), a few sites in the Pilbara and Carnarvon Basin (Halse et al. 2000; Pinder et al. 2010), and Hutt Lagoon (Pinder et al. 2012), but not south of this. This species was not collected during the 1999 survey. The Brachionus (plicatilis group) collected from Eil Eil Spring and the main Stockyard Spring belongs to a cryptic species complex. The group is very difficult to identify using morphological characters, so DNA taxonomy has been used to sleuth this group further (Mills 2016; Mills et al. 2016). Rotifers tend to be widespread so this species is unlikely to be locally endemic.

Acarina A potentially new water mite, Arrenurus (Micruracarus) MAN1, was collected in 2015 from four sites including the outer pools at Fern Spring, Little Eil Eil Spring, Eil Eil Spring, and Bretts Spring. This does not resemble any published species or any other species in the Parks and Wildlife collection.

Crustacea Cladocera: A number of specimens from the family Chydoridae collected during the survey are still being reviewed by taxonomic experts. At present, it is likely that cf. Celsinotum n.sp (collected from sites 6 and 7B) is new (R Shiel, pers. comm., 2016). The remaining cladocerans collected are relatively common and widespread. Copepoda: Calanoid copepods were noticeably absent from all wetlands during the survey. The cyclopoid copepod Halicyclops spinifera, recorded from several of the Walyarta mounds springs (sites 3, 4 and 5) in 2015 and from Little Eil Eil in 1999, is relatively uncommon. In WA, this species has otherwise been collected from Lake Thetis near Jurien Bay (Pinder & Quinlan 2015) and from groundwater in Cape Range (Pesce et al. 1996). The remaining cyclopoids are all fairly common and widespread. Some harpacticoid species belong to genera with marine origins and tend to be associated with brackish to saline waters with connections to the coast. Two of these, Onychocamptus bengalensis and Cletocamptus dietersi are cosmopolitan species collected from a number of the Walyarta springs and the former also from Salt Creek. These have been commonly collected in Western Australia. Robertsonia propinqua is a circum-tropical species inhabiting marine littoral, estuarine and saline wetlands. It occurred in both of the Salt Creek sites (18 and 19) during the most recent survey but has not been commonly collected in Western Australia: Lake MacLeod during a survey of the Carnarvon Basin (Halse et al. 2000) and from Wheatbelt salt lakes (Bayly & Williams 1966; Hamond 1973). As noted in the groundwater

30 section above, one of the three undescribed Schizopera collected at Walyarta has not been seen from elsewhere (J McRae, pers. comm., 2016). This species is probably stygophilic and was collected from Saunders Spring (5) and Little Eil Eil Spring (6) in 2015, but no Schizopera were collected in 1999. Ostracoda: It is thought that a cypridid ostracod collected from Fern, Melaleuca, and Eil Eil springs (sites 3B, 4 and 7A) is new (S Halse, pers. comm., 2016). It is currently being reviewed and formally described by taxonomic experts, but has been listed in Appendix 2 as Cyprididae n. gen. n.sp. The Heterocypris collected from sites 5, 9, 12 and 23 is also potentially new. Molecular work has been undertaken on this group, but the wider distribution of this species is not yet known (S Halse, pers. comm., 2016). It is listed in Appendix 2 as Heterocypris sp. MAN1 (nr vatia). Syncarida: Storey et al. (2011) collected specimens of a then new species of syncarid: Kimberleybathynella mandorana Cho et al. 2005. This was collected from the interstitial sample from Fern Spring but was not collected in 2015. Insecta

Coleoptera: A Haliplus water beetle collected from Eil Eil Spring is a potentially new species (C Watts, pers. comm. 2016). The specimens from Walyarta do not currently match any formally described species of Haliplus. It does share some affinity with H. bistriatus which occurs mostly in eastern Australia, but the Mandora species differs in colouration and lacks a diagnostic character of H. bistriatus. Agraphydrus coomani has rarely been collected in Western Australia, though it is known from the Kimberley region and a few Pilbara records (Atlas of Living Australia, accessed 03 Nov 2016) but it was not collected during the Pilbara Biological Survey. Chasmogenus nitescens appears not to have been collected in Western Australia previously but it is otherwise widely distributed in northern Australia and the southwest Pacific. None of these three beetles were collected in 1999. Hemiptera: The collection of one of the small water striders, Microvelia (Pacificovelia) lilliput (Veliidae), from Little Eil Eil Spring and Eil Eil Spring pools (sites 6 and 7B) was a significant record. Previously known from more northern parts of Australia, this record from Walyarta may be the most southern record for this species in WA, (T Weir, pers. comm., 2016). No veliids were collected in 1999. Gastropoda Assimineid snails were collected from six springs in 2015. No comparisons have been made with the Assiminea sp. nov. collected in 1999 from Fern Spring and Linear Spring but it is almost certainly the same species. This snail was originally collected from along the water’s edge at Eil Eil spring by Ponder (unpubl. data), with related species found in the Kimberley and in a few springs in the Northern Territory.

31 Fish During the 1999 survey, a new species of goby (Acentrogobius sp. nov.) was collected from the Salt Creek at Walyarta (Storey et al. 2011). In 2015, additional specimens were collected from this same area and handed over to the Museum and Art Gallery of the Northern Territory. These new specimens will aid genetic work and contribute to the formal description of this new species.

32 5. Comparison between the present survey and the 1999 survey

A total of 134 aquatic invertebrate taxa were recorded from 10 sites in 1999, compared to 175 taxa collected from 11 sites in 2015. Eight of the sites sampled in 2015 had previously been sampled for invertebrates by Storey et al. (2011) in 1999, and it is only these sites which are considered further in this section. A comparison of species richness for each site between years is provided in Table 4. At four of the springs (5, 6, 7 and 9), richness in 2015 is about double (or more) what it was in 1999, although a plankton sample was not collected at Top Spring (site 9) in 1999. An average of 27 taxa was collected per site (excluding site 9) in 1999, compared to an average of 47 taxa per site in 2015. Of a combined total of ~200 taxa (from these eight sites), just under a third were recorded in both years, with 101 taxa recorded only in 2015, and 41 recorded only in 1999. It should be noted that species richness varies slightly between the 2015 data (detailed in section 4.2 above) and the combined (1999 & 2015) dataset (below). This is due to the need to standardise taxonomic resolution across years. For example, all genera belonging to the family Ceratopogonidae (Diptera) collected in 2015 (i.e. Bezzia, Culicoides, Dasyhelea, Forcipomyia, Monohelea, and Nilobezzia) have been grouped together as Ceratopogonidae for this comparison, so that identifications are consistent with Storey et al. (2011).

Table 4: Species richness at eight sites sampled at Walyarta in both August 1999 and September 2015. Site No. Site Name 1999 2015 3 Fern Spring 31 54 4 Melaleuca Spring 20 30 5 Saunders Spring 21 41 6 Little Eil Eil Spring 32 62 7 Eil Eil Spring 36 82 9 Top Spring 9 58 17 Bretts Spring 36 38 18 Salt Creek Spring 15 16

An nMDS ordination of all sites from Walyarta that were sampled in both 1999 and 2015 (excluding site 9) is shown in Fig. 14, with data from subsites (e.g. sites 3 and 7) combined in this ordination. The ordination showed a separation of invertebrate community composition between years (i.e. species composition at a particular site in 1999 is quite dissimilar to the composition at the same site in 2015). The associated ANOSIM analysis showed a significant difference between years (ANOSIM; Global R = 0.548, p < 0.001).

33 2D Stress: 0.12 Year 6 1999 2015 7 3 5

4

17 7 6

5 4 17 18

3

18

Figure 14: Two-dimensional nMDS ordination of invertebrate communities in 7 wetlands at Walyarta, sampled in both 1999 (blue) and 2015 (red).

20 YEAR 1999 2015

40 A B C D E F y t i r a l i 60 m i S

80

100 9 5 5 5 5 5 5 5 9 9 9 9 9 9 9 1 1 1 1 1 1 1 9 9 9 9 9 9 ------8 8 6 7 5 7 3 4 7 6 7 4 3 5 1 1 1 1 Samples

Figure 15: Cluster analysis (UPGMA) of invertebrate communities in 7 wetlands at Walyarta, sampled in 1999 and 2015. Black lines show divisions which have statistical support, and red lines indicate sub- structure with no statistical support.

34

A hierarchical group-average cluster analysis (Fig. 15) shows two distinct clusters of 1999 (groups D, E, F) and 2015 sites (groups B & C), with the exception of site 18 (Salt Creek Spring, group A). In both 1999 and 2015 sites 6 and 7 (Eil Eil and Little Eil Eil) grouped together (groups B & D), as did sites 3, 4 and 5 (groups C & F). Bretts Spring (17) clustered with sites 3, 4 and 5 in both years, but was more differentiated from these sites in 1999 (as group E). This clustering trend can also be observed in the ordination in Figure 14. This analysis shows that while invertebrate community composition at sites in 1999 was quite dissimilar to that recorded in 2015, each of the same wetlands tended to group together in both years.

A SIMPER analysis was used to understand which invertebrate taxa were contributing most to the dissimilarity observed between these groups. A comparison of groups C and F (primarily sites 3, 4 and 5 in 2015 and 1999 respectively) revealed nine taxa that were contributing most to this dissimilarity. These were flatworms, beetles (Hydroglyphus leai, Hydaticus consanguineus and Regimbartia attenuata), Anopheles mosquitoes, the chironomid Procladius paludicola, and dragonflies/damselflies (Ischnura aurora aurora, Diplacodes bipunctata\trivialis, and Orthetrum caledonicum). Of these, the beetles were all recorded in 1999 and not 2015, and the remaining groups/species were all recorded in 2015 and not in 1999. A further comparison, this time of the Little Eil Eil and Eil Eil suite between years (groups B & D) revealed that 45% of the variation observed between these sites is attributed to more than 30 species. It is clear from these analyses that there was a major difference in invertebrate composition between 1999 and 2015 but that patterns in aquatic invertebrate occurrence across the springs was quite similar. This indicates a high turnover of species but it is not clear over what time periods this occurs (because we have only sampled twice) or what is driving the difference. There is very little information available on species turnover in arid-zone wetlands, so it is unclear whether the level of turnover between surveys is to be expected. The average percentage of species in common between both years per site ranged from 10% of species in common (sites 5 and 6) to 36.4% (site 18). These differences are discussed in more detail below for each major taxonomic group:- Protists and Rotifers The number of protists and rotifers was similar between years, although the species collected from each year did vary. Species from the family Lepadellidae were collected in 2015 but not in 1999. Oligochaeta The number and diversity of oligochaetes was similar between years, although specimens from the family Enchytraeidae were additionally collected in 2015. Acarina Seven taxa of water mites were collected in 2015 compared with only three species in 1999. Arrenurus mites were collected in both years but it is not clear whether the unidentified specimens collected in 1999 are the potentially new Arrenurus water mite collected in 2015.

35

Cladocera Noticeably absent from many sites in both 1999 and 2015 were cladocerans, with only two species recorded in 1999 (site 7 and 17), and seven taxa from four of the eight sites (6, 7, 9 and 17) in 2015. Cladocerans tend to be more diverse in larger open water wetlands.

Copepoda The diversity of copepods varied slightly between years, with seven species collected in 1999 and 11 in 2015, with four species collected in both years (Halicyclops spinifera, Mesocyclops brooksi, Cletocamptus dietersi and Onychocamptus bengalensis). A major difference is the absence of Schizopera species in 1999 compared to three present in 2015.

Ostracoda All species collected from these eight sites in 1999 were also recorded in 2015. Two genera, Heterocypris and Penthesilenula, were collected in 2015 but not in 1999, together with the new cypridid ostracod from Fern, Melaleuca and Eil Eil Springs.

Coleoptera Beetles from the genera Coelostoma, Copelatus, Cybister, Hydrochus, Regimbartia and Sternopriscus were all recorded in 1999 but absent in 2015. Beetles from the genera Agraphydrus, Chasmogenus, Dineutus and Limbodessus were all collected in 2015 but were absent in 1999. Beetles are highly mobile insects and these differences indicate substantial immigration and emigration over time.

Hemiptera Nine hemipterans were recorded in 1999 and fourteen in 2015, with four species collected in both years (Diplonychus eques, Agraptocorixa parvipunctata, Micronecta virgata, Anisops nasutus). Genera that were collected in 2015 but absent in 1999 were Microvelia, Mesovelia, Hebrus and gerrids.

Odonata No damselflies and only two dragonfly species (Orthetrum caledonicum, Tramea stenoloba/loewii) were collected across the eight sites sampled in 1999, although one damselfly was collected at Coolabah Claypan (not considered here). By comparison, at these same eight sites, two species of damselflies and seven species of dragonflies were collected in 2015. Again, this is a highly mobile group of insects (especially the dragonflies).

Trichoptera In 1999 Triplectides ciuskus seductus was recorded from only one of the same eight sites being compared (Bretts Spring), although Oecetis and Ecnomus were recorded in 1999 from Coolabah Claypan, whereas in 2015 species of Oecetis and Triplectides were collected from several of the springs, but none were collected from Bretts Spring.

36 Diptera The level of taxonomic resolution for this group varied considerably between 1999 and 2015, making direct comparisons between species lists difficult. Some groups (i.e. Ceratopogonidae) were left at a broader taxonomic resolution (i.e. family level) in 1999 but were resolved further to Bezzia, Culicoides, Monohelea, Nilobezzia, Dasyheleinae and Forcipomyiinae in 2015.

Other taxa Diversity of molluscs and mayflies was similar between years. Assimineid snails were collected from six springs in 2015. No comparisons have been made with the Assiminea sp. nov collected in 1999, but it is almost certainly the same species. A locally endemic syncarid, Kimberleybathynella mandorana, collected from an interstitial sample at Fern Spring 1999 and described by Cho et al. (2005) was not collected during the 2015 survey.

Why the difference? Wetlands, especially those that have only temporary presence of water, are highly dynamic ecosystems, with communities changing within and between years in response to physical and chemical changes in conditions as well as ecological and biological processes including colonisation, trophic dynamics and life-cycle traits (Williams 2006). In arid zones, springs are particularly important refuges from drought, providing a more reliable source of moisture to enable survival during drier times (Brim-Box et al. 2008). Nonetheless, even springs can be quite dynamic systems. The Mandora springs provide two main types of aquatic habitats, a mound of peat, often with small pools of shallow water and an outer pool or moat fed by seepage from the mound. In 2015, some of the mounds (such as Fern Spring) were entirely wet and boggy at the surface and had small pools while others (such as Grants Spring) were largely dry but still have an outer pool. At least some of the outer pools are not permanent, but we do not know how frequently these flood and dry. Figure 16 shows Eil Eil outer pool with water on 8 Sep 2015 (3 days after it was sampled for this project) and two weeks later on 21 Sep 2015. Some drying and flooding may be in response to tidal movements of groundwater influencing discharge (Jasmine Rutherford, DPaW, pers. comm.), so flooding and drying may occur regularly and be partly uncoupled from rainfall.

Communities in temporary waters start to develop within days of first inundation and continue to develop for several months as species hatch from drought resistant life-stages or immigrate from elsewhere (including as parasites such as water mites or via waterbirds) (Boulton and Lloyd 1992; Brock et al. 2003; Brooks 2000; Siziba et al, 2013). In the Walyarta mound springs there would also be some species that are able to survive in moist habitats within or on the peat mounds or in groundwater, as evidenced by invertebrates in the interstitial samples. If the outer pools fluctuate with tidal movements of groundwater then some species must be able to tolerate periodic (and perhaps quite frequent) absence of surface water. In two temporary wetlands in the Wheatbelt sampled by Pinder et al. (2013) on 8 occasions over a 5 month inundation, species richness on any one sampling occasion was only half (or less) of the species

37 present across the season, peaking after 3-4 months. This shows that samples taken in different years can be quite different if they are taken at different stages on the hydrological cycle. In 2015, the diversity in our samples, and the fact that few species were present only as early developmental stages, suggests we sampled after the pools had been around for sufficient time for a full community to develop. Overlain on top of the seasonal (or aseasonal) cycles of filling and drying of individual pools in arid zones are major differences in rainfall between years (and multiple years). Multiple wet years may allow some fauna to arrive and survive that would not remain during consecutive dry years.

Figure 16: Photographs of Eil Eil Outer Pool (7A) taken A) on 8 Sep 2015 three days after it was sampled and B) two weeks after it was sampled. Photos by Shane Sercombe (2-Mile Contracting).

The marsh and surrounds were extensively flooded in 1999 and had experienced higher than average rainfall in the five years preceding the survey, enabling sampling of wetlands which were dry in 2015, such as Lake Walyarta and Coolibah Claypan in 1999. This presents an enigma: Why were more species collected in 2015 (a dry year following a previously dry year) than in 1999 (a wet year following several wet years that should have promoted diversity). This is not a simple question to answer. Large flood events over a number of years may have partially reset the fauna at some of the springs by washing out some of the pools. Also, the area covered by surface water in 1999 (and preceding years) was presumably larger than in 2015 which may have meant that the fauna were less concentrated on a few refuges than when smaller areas of wetland remain.

There are no obvious differences in water chemistry between 1999 and 2015 that would account for the differences in richness and composition of the aquatic invertebrate communities. Total nitrogen concentrations were higher in 1999 compared to 2015, with the exception of Bretts Spring. However, while Melaleuca Spring had particularly high nutrient concentrations (17 mg.L-1 total N and 2 mg.L-1 total P) in 1999 than in 2015 (0.68 mg.L-1 and 0.1

38 mg.L-1) the difference in invertebrate richness between 1999 and 2015 was lower than for most other springs (Table 3). Other environmental variables were reasonably similar between years. An nMDS ordination of environmental data from both years did not reveal a strong grouping of sites according to year and there was no significant differences in overall water quality between years (ANOSIM; Global R=0.071, p > 0.1).

Sampling methods appear to have been comparable in the two years, with sweep net samples of up to 50 metres in length taken (Storey et al. 2011). With a few exceptions taxonomic resolution within the dataset was also comparable between the two years. Nonetheless, the difficulty in sampling shallow springs and the potential for under-estimating aquatic invertebrate species richness was acknowledged by Storey et al. (2011) and this may account for some of the variation in the fauna collected on each of the sampling occasions.

Rather than comparing the 1999 and 2015 data to investigate differences in the fauna over 16 years, the two datasets should be taken as complementary, together presenting a fuller picture of the aquatic invertebrate diversity that utilises these dynamic arid zone wetlands over the range of climatic and hydrological conditions they experience.

39 References

ANZECC & ARMCANZ (2000) Australian and New Zealand Guidelines for Fresh and Marine Waters. National Water Quality Management Strategy Paper No 4, Australian and New Zealand Environment and Conservation Council & Agriculture Resource Management Council of Australia and New Zealand, Canberra. Bayly IAE, Williams WD (1966) Chemical and biological studies on some saline lakes of south- east Australia. Australian Journal of Marine and Freshwater Research 17, 177–228. BirdLife International (2016). Important Bird Areas Factsheet: Mandora Marsh and Anna Plains. Downloaded from http://www.birdlife.org August 2016. Boulton AJ, Lloyd LN (1991) Macroinvertebrate assemblages in floodplain habitats of the lower River Murray, South Australia. Regulated Rivers Research and Management 6, 183–201. Boulton AJ, Lloyd LN (1992) Flooding frequency and invertebrate emergence from dry floodplain sediments of the River Murray. Regulated Rivers Research and Management 7, 137–151. Boulton AJ, Sheldon F, Jenkins KH (2006) Natural disturbance and aquatic invertebrates in desert rivers. In: Kingsford RT (ed) Ecology of desert rivers. Academic, New York, 133-153 pp. Bray, JR, Curtis JT (1957). An ordination of upland forest communities of southern Wisconsin. Ecological Monographs 27, 325–349. Brim-Box J, Duguid A, Read RE, Kimber RG, Knapton A, Davis J, (2008) Central Australian waterbodies: The importance of permanence in a desert landscape. Journal of Arid Environments 72, 1395–1413. Brock MA, Nielsen DL, Shiel RJ, Green JD, Langley JD (2003) Drought and aquatic community resilience: the role of eggs and seeds in sediments of temporary wetlands. Freshwater Biology 48, 1207–1218. Brooks R (2000) Annual and seasonal variation and the effects of hydroperiod on benthic macroinvertebrates of seasonal forest (“vernal”) ponds in central Massachusetts, USA. Wetlands 20, 707–715. Bureau of Meteorology (2016) Climate data online, downloaded from http://www.bom.gov.au/climate/data/index.shtml Accessed August 2016. Cho J-L, Park J-G, Humphreys WF (2005) A new genus and six new species of the Parabathynellidae (Bathynellacea, Syncarida) from the Kimberley region, Western Australia. Journal of Natural History 39(24), 2225–2255. Clarke KR, Gorley RN (2015). Getting started with PRIMER v7: User Manual/Tutorial. PRIMER-E Ltd, Plymouth U.K.

40 Clarke KR, Somerfield PJ, Gorley RN (2008) Testing of null hypotheses in exploratory community analyses: similarity profiles and biota-environment linkage. Journal of Experimental Marine Biology and Ecology 366, 56–69. Daniel G, Kern S, Pinder A, Nowicki A (2009) Resource Condition Report for a Significant Western Australian Wetland: Saunders Spring. Prepared for Inland Aquatic Integrity Resource Condition Monitoring (IAI RCM) Project. Department of Environment and Conservation. Perth, Australia. English V, Luu R, Coote M (2016) Survey of Mandora Mound Springs and Salt Creek, Walyarta. Unpublished report, Department of Parks and Wildlife, Kensington. 26 pp. Environment Australia (2016) Directory of Important Wetlands in Australia - Information sheet: Mandora Salt Marsh-WA042. Downloaded from http://www.environment.gov.au August 2016. Graham G (1999) A Land Management Assessment of Mandora Marsh / Walyarta and Its Immediate Surrounds. Unpublished report to Environment Australia, Department of Conservation and Land Management, Kununurra, Western Australia. 91 pp. Hale J, Butcher R (2009) Ecological Character Description of the Eighty-mile Beach Ramsar Site. Report to the Department of Environment and Conservation, Perth, Western Australia. Halse SA, Scanlon MD, Cocking JS (2002) Do springs provide a window to the groundwater fauna of the Australian arid zone. In: Balancing the Groundwater Budget: Proceedings of an International Groundwater Conference, Darwin. Halse SA, Shiel RJ, Storey AW, Edward DHD, Lansbury I, Cale DJ, Harvey MS (2000) Aquatic invertebrates and waterbirds of wetlands and rivers of the southern Carnarvon Basin, Western Australia. Records of the Western Australian Museum Supplement 61, 217–26. Halse SA, Pearson GB, Hassell C, Collins P, Scanlon MD, Minton CDT (2005) Mandora Marsh, north-western Australia, an arid-zone wetland in maintaining continental populations of waterbirds. Emu 105, 115–125. Hamond R (1973) The Australian species of Robertsonia (Crustacea, Harpacticoida), with a revised key to the genus. Records of the Australian Museum 28, 421–435. Johnstone RE, Burbidge AH, JC Darnell JC (2013) Birds of the Pilbara region, including seas and offshore islands, Western Australia: distribution, status and historical changes. Records of the Western Australian Museum Supplement 78, 343–441. Karanovic T (2006) Subterranean copepods (Crustacea, Copepoda) from the Pilbara region in Western Australia. Records of the Western Australian Museum Supplement 70, 1–239. Karanovic T, Cooper SJB (2012) Explosive radiation of the genus Schizopera on a small subterranean island in Western Australia (Copepoda: Harpacticoida): unravelling the cases of cryptic speciation, size differentiation and multiple invasions. Invertebrate Systematics 26, 115– 192.

41 Karanovic T, McRae J (2013) The genus Schizopera (Copepoda, Harpacticoida) in the Pilbara region of Western Australia, with description of a new species and its molecular and morphological affinities. Records of the Western Australian Museum 28, 119–140. Mills S (2006) Investigations of the Brachionus plicatilis species complex, with particular reference to southwest Western Australia. Ph.D. thesis, University of Western Australia, Perth. Mills S, Alca´ntara-Rodrı´guez JA, Ciros-Pe´rez J, Go´mez A, Hagiwara A, Hinson Galindo K, Jersabek CD, Malekzadeh-Viayeh R, Leasi F, Lee J-S, Mark Welch DB, Papakostas S, Riss S, Segers H, Serra M, Shiel R, Smolak R, Snell TW, Stelzer CP, Tang CQ, Wallace RL, Fontaneto D, Walsh EJ (2016) Fifteen species in one: deciphering the Brachionus plicatilis species complex (Rotifera, Monogononta) through DNA taxonomy. Hydrobiologia doi: 10.1007/s10750-016-2725-7. Pesce GL, de Laurentiis P, Humphreys WF (1996) Copepods from ground waters of Western Australia. II. The genus Halicyclops (Crustacea, Copepoda, Cyclopidae). Records of the Western Australian Museum 18, 77–85. Pinder AM, Halse SA, McRae JM, Shiel RJ (2004) Aquatic invertebrate assemblages of wetlands and rivers in the Wheatbelt region of Western Australia. Records of the Western Australian Museum Supplement 67, 7–37. Pinder AM, Halse SA, Shiel RJ, McRae JM (2010) An arid zone awash with diversity: patterns in the distribution of aquatic invertebrates in the Pilbara region of Western Australia. Records of the Western Australian Museum Supplement 78, 205–246. Pinder AM, Leung AE (2010) Report on the August 2008 sampling of the aquatic invertebrate assemblages of mound springs of the Three Springs Area Threatened Ecological Community. Department of Environment and Conservation, Perth. Pinder AM, Quinlan K (2015) Aquatic invertebrate communities of wetlands along the Jurien coast of Western Australia. Journal of the Royal Society of Western Australia 98, 69–88. Pinder AM, Quinlan K, Cale DJ, Leung AE (2012) Aquatic invertebrates of the Hutt River and Hutt Lagoon catchments, Western Australia. Journal of the Royal Society of Western Australia 95, 29– 51. Pinder AM, Quinlan K, Cale DJ, Shiel RJ (2013) Invertebrate community development over a hydrological cycle in seasonal claypans of Drummond Nature Reserve, Western Australia (with updated cladoceran data in January 2015). Unpublished report, Department of Parks and Wildlife, Kensington. Ponder WF (unpubl data) Report on a preliminary survey of springs in the Pilbara Region of Western Australia. Australian Museum, Sydney, 17 pp. Segers H (2007) Annotated checklist of the rotifers (Phylum Rotifera), with notes on nomenclature, taxonomy and distribution. Zootaxa 1564, 1–104.

42 Semeniuk C, Semeniuk V (2000) Wetlands of the north western Great Sandy Desert in the LaGrange hydrological sub-basin. Unpublished report for the Water and Rivers Commission, Perth. Sheldon F, Boulton AJ, Puckridge JT (2002) Conservation value of variable habitat connectivity: aquatic invertebrate assemblages of channel and floodplain habitats of a central Australian arid- zone river, Cooper Creek. Biological Conservation 103, 13–31. Siziba N, Chimbari MJ, Mosepele K, Masundire H, Ramberg L (2013) Inundation frequency and viability of microcrustacean propagules in soils of temporary aquatic habitats of lower Okavango Delta, Botswana. Ecohydrology 6, 722–730. Start AN, Kendrick P, McKenzie NL (2008) Biota of the Mandora System, Western Australia: - Mammals. Journal of the Royal Society of Western Australia 91, 209–217. Storey AW, Halse SA, Shiel RJ, Creagh S (2011) Aquatic fauna and water chemistry of the mound springs and wetlands of Mandora Marsh, north-western Australia. Journal of the Royal Society of Western Australia 94, 419-437. Williams DD (2006) The Biology of Temporary Waters. Oxford University Press, Oxford. Wollheim WM, Lovvorn JR (1996) Effects of macrophyte growth forms on invertebrate communities in saline lakes of the Wyoming High Plains. Hydrobiologia 323, 83–96.

43 Appendix 1 – Water quality parameters and habitat information for springs and wetlands surveyed at Walyarta in 2015.

Water Chemistry - Field Water Chemistry - Laboratory Site Water Site Name Code Temp Conductivity Salinity pH Turbidity ECond Alkalinity Colour Na Ca K Mg Cl HCO3 CO3 SO4 °C uS cm-1 g L -1 NTU mS/m mg L-1 TCU %meqL-1 %meqL-1 %meqL-1 %meqL-1 %meqL-1 %meqL-1 %meqL-1 %meqL-1 Fern Spring inner pool 3A 22.2 13570 7.4 7.71 1 1310 165 130 82.32 10.14 1.96 5.58 83.01 2.49 0.01 14.48 Fern Spring outer pools 3B 22.2 13490 7.4 7.71 0.25 1300 199 140 82.31 10.07 1.84 5.78 82.26 2.94 0.01 14.78 Melaleuca Spring 4 28.9 11640 6.28 8.59 4.1 1180 200 230 85.24 8.95 1.16 4.66 84.47 3.09 0.22 12.22 Saunders Spring 5 28.4 3740 1.94 8.66 49 312 351 280 81.93 10.91 1.80 5.37 67.24 23.10 0.05 9.60 Little Eil Eil Spring 6 20 2310 1.171 7.4 0.7 230 194 150 80.33 12.50 1.39 5.78 73.38 17.89 0.08 8.65 Eil Eil Spring 7A 34.4 8340 4.39 8.25 13 829 408 80 85.42 7.11 1.61 5.85 76.78 9.86 0.02 13.34 Eil Eil Spring - pools 7B 21.7 3750 1.74 8.31 ------Top Spring 9 25.4 2610 1.23 8.66 0.6 141 313 100 76.42 13.41 2.87 7.29 46.26 44.11 0.12 9.52 Main Stockyard spring 12 22.8 4950 2.57 8.75 8.1 471 150 36 81.00 10.91 2.35 5.74 78.52 6.96 0.04 14.48 Small mangrove spring 13 22.4 3490 1.8 7.69 ------Spring 200m ESE of stockyard 14 23.4 4130 2.12 8.56 ------Spring 100m W of stockyard 15 23.3 7050 3.73 8.46 ------Spring 150m W of stockyard 16 21.2 3580 1.84 6.63 ------Bretts Spring 17 26.1 8110 4.32 8.23 5.3 797 348 70 87.53 5.95 1.47 5.05 80.09 9.27 0.18 10.46 Salt Creek Spring 18 24 73600 35.7 8.08 0.25 4680 234 5 86.94 2.59 5.80 4.67 80.74 0.90 0.03 18.32 Salt Creek Stromatolite Pool 19 24.3 61900 37.1 8.99 1 6290 178 9 91.67 1.33 4.05 2.96 82.81 0.31 0.20 16.68 Grants Spring 23 31.1 4690 2.35 8.25 170 438 495 450 86.37 8.11 1.85 3.67 70.40 24.01 0.04 5.55

44

Appendix 1 – continued.

Nutrients Habitat Information Submerged Emergent Site Name TN TFN TP TFP Chlor-a Phaeo-a Sample Depth Stones Sand Silt Clay macro cover macro cover Site Code mg L-1 mg L-1 ug L-1 ug L-1 ug L-1 ug L-1 cm % % % % % % Fern Spring inner pool 3A 1.2 0.82 49 16 2 0.5 20 0.8 84.5 4 11.5 0 5 Fern Spring outer pools 3B 0.40 0.34 5 5 0.5 0.5 20 0.05 31 34 35 2 0 Melaleuca Spring 4 0.68 0.53 100 100 10 2 10 0.05 50 30 20 60 0 Saunders Spring 5 2.7 1.7 730 400 7 2 15 0.05 37.5 40 22.5 70 0 Little Eil Eil Spring 6 0.35 0.24 49 14 0.5 0.5 25 0.7 48 28 24 50 0 Eil Eil Spring 7A 2.2 1.6 230 98 2 0.5 19 0.05 53.5 37.5 9 60 5 Eil Eil Spring - pools 7B ------18 0.4 52.5 29.5 18 90 2 Top Spring 9 0.78 0.60 29 18 2 2 15 1.4 51.5 30.5 18 50 1 Main Stockyard spring 12 1.8 0.75 190 35 4 0.5 20 3.1 63 27 10 1 1 Small mangrove spring 13 ------Spring 200m ESE of stockyard 14 ------Spring 100m W of stockyard 15 ------Spring 150m W of stockyard 16 ------Bretts Spring 17 1.3 0.97 72 43 3 0.5 19 0.05 47.5 40 12.5 10 25 Salt Creek Spring 18 0.99 0.83 5 5 0.5 0.5 45 2.7 85.5 6 8.5 5 0 Salt Creek Stromatolite Pool 19 1.4 1.2 5 5 0.5 0.5 51 0.05 40.5 35 24.5 0 10 Grants Spring 23 4.6 2.3 540 73 8 1 40 0.05 48.5 33 18.5 2 25

45 Appendix 2 – Aquatic Invertebrate Species List (2015) At least 175 distinct species of aquatic invertebrates were collected from 11 wetlands at Walyarta in September 2015. 203 taxa are listed below, as some of these (*) likely represent a species already listed, but are just identified at a different level of taxonomic resolution. (INT) refers to an interstitial sample.

3A 3A 3B 4 4 5 5 6 6 7A 7B 7B 9 12 17 18 19 23 23 Lowest Identification (INT) (INT) (INT) (INT) (INT) (INT)

PROTISTA Arcella discoides 1 1 Arcella hemisphaerica 1 Arcella bathystoma 1 Arcella sp. a 1 1 Arcella sp. b 1 1 Centropyxis aculeata 1 1 Centropyxis ecornis 1 Centropyxis platystoma 1 Centropyxis sp.* 1 Euglypha sp. 1

- Turbellaria 1 1 1 1 1 1 1 1 - Microturbellaria 1 - Nematoda 1 1 1 1 1 1 1 1 1 1 1 1

ROT IFERA Rotaria sp. 1 Bdelloidea* 1 1 1 Brachionus angularis 1 Brachionus quadridentatus 1 Brachionus (plicatilis group) 1 1 Keratella australis 1 Colurella cf. obtusa 1 Lepadella patella 1

46 Lepadella sp. a 1 Lecane bulla 1 1 Lecane grandis 1 1 Lecane luna 1 Lecane papuana 1 Lecane signifera 1 Lecane stenroosi 1 1 1 Lecane ungulata 1 Lecane sp. s.str.* 1 Cephalodella forficula 1 Synchaeta sp. 1

GASTROPODA Gyraulus sp. 1 1 1 1 1 1 1 1 1 Assiminaeidae 1 1 1 1 1 1 1 1 1 1

OLIGOCHAETA Naididae (ex Tubificidae) 1 Dero furcata 1 1 1 1 Allonais ranauana 1 1 Pristina longiseta 1 1 1 1 1 Enchytraeidae MAN1 1 1 1 1 1 Enchytraeidae MAN2 1 Enchytraeidae MAN3 1

ACARINA Hydrachna sp. 1 Hydrodroma sp. 1 Neumania sp. 1 Arrenurus balladoniensis 1 1 1 Arrenurus sp.* 1 Arrenurus (Micruracarus) MAN1 1 1 1 1 1 Acarina* 1 Oribatida sp.* 1 1 1 Mesostigmata 1 Trombidioidea 1

47 Oribatida type 2 1 1 Oribatida type 1 1 1 1 1

CLADOCERA Latonopsis australis 1 Alonella sp. 1 cf. Celsinotum sp. a 1 cf. Celsinotum n. sp 1 1 Dunhevedia crassa 1 1 Leberis diaphanus 1 1 1 1 1 Anthalona harti 1 Chydoridae* 1 Ceriodaphnia cornuta 1 1 1 Ceriodaphnia sp.* 1 1 1 1 Macrothrix sp. 1

OSTRACODA Penthesilenula brasiliensis 1 Vestalenula marmonieri 1 1 Vestalenula matildae 1 Darwinulidae* 1 1 Candonopsis tenuis 1 1 1 1 Candonopsis sp.* 1 Cypretta sp. BOS609 1 Cypretta sp. BOS610 1 1 1 Cypretta sp. BOS611 1 Cyprinotus cingalensis 1 Diacypris spinosa 1 1 Heterocypris sp.* 1 Heterocypris sp. MAN1 (nr vatia) 1 1 1 1 Cypricercus sp. 1 Cyprididae n. gen. n.sp. 1 1 1 Cypridopsis sp. 1

COPEPODA Microcyclops varicans 1 1 1

48 Metacyclops aff mortoni 1 Halicyclops spinifer 1 1 1 Ectocyclops phaleratus 1 1 1 Mesocyclops brooksi 1 1 1 1 1 1 1 1 1 1 1 Paracyclops nr chiltoni 1 Apocyclops dengizicus 1 1 Cyclopoida* 1 1 Cletocamptus deitersi 1 1 1 1 1 1 1 1 1 Onychocamptus bengalensis 1 1 1 1 1 Robertsonia propinqua 1 1 Schizopera sp. B22 1 Schizopera sp. B23 1 1 Schizopera sp. B24 1 1 Nitokra sp.* 1 Nitokra lacustris sp. B04 1 Nitokra lacustris sp. B05 1 1 Harpacticoida* 1

COLEOPTERA Haliplus sp. (nr bistriatus) 1 Haliplidae (larva)* 1 Laccophilus sharpi 1 Hyphydrus elegans 1 Hyphydrus lyratus 1 1 Hyphydrus sp. (larva)* 1 1 1 1 Hydroglyphus trifasciatus 1 1 1 1 Hydroglyphus grammopterus 1 1 Limbodessus compactus 1 Allodessus bistrigatus 1 1 1 1 1 1 1 1 Platynectes sp. (larva) 1 Rhantus sp. (larva) 1 Eretes australis 1 1 1 1 Hydaticus sp. (larva) 1 Dineutus (Cyclous) australis 1 1 1

49 Berosus australiae 1 1 1 1 1 Berosus approximans 1 Berosus dallasae 1 Berosus pulchellus 1 1 1 1 Berosus sp. (larva)* 1 1 1 1 1 1 1 1 1 1 Enochrus (Methydrus) elongatulus 1 Enochrus (Methydrus) deserticola 1 1 1 Enochrus fuscatus (was malabarensis) 1 1 1 1 1 1 1 1 1 Enochrus sp. (larva)* 1 1 Helochares tatei 1 Helochares sp. (larva)* 1 1 1 1 Paracymus sp. 1 1 Sternolophus sp. (larva) 1 1 1 1 Chasmogenus nitescens 1 Agraphydrus (Agraphydrus) coomani 1 1 1 Hydrophilidae* 1 Ochthebius nr sp. 4 1 1 1 1 1 1 1 Scirtidae 1 1 1

DIPTERA Tipulidae type A 1 Tipulidae type E 1 Anopheles spp. 1 1 1 1 1 1 1 1 1 1 1 1 Aedes sp. 1 Culex (Culex) annulirostris 1 1 1 1 1 1 1 1 1 Culex (Oculeomyia) bitaeniorhynchus 1 Culex (Oculeomyia) starckeae 1 1 Culex sp.* 1 Bezzia sp. 1 1 1 1 1 1 1 1 Culicoides sp. 1 1 1 1 1 1 1 1 1 Monohelea spp. 1 1 1 1 1 1 1 1 1 1 1 Nilobezzia sp. 1 1 1 1 1 1 1 1 1 Dasyheleinae 1 1 1 1 1 1 1 1 1 1 1 1 1 Forcypomyinae 1 1 1

50 Psychodinae sp. 3 1 Psychodinae sp. 5 1 Scatopsidae 1 Tabanidae 1 1 1 1 Stratiomyidae 1 1 1 1 1 1 1 1 1 1 1 1 1 Dolichopodidae 1 1 1 Syrphidae 1 1 1 Ephydridae spp. 1 1 1 1 1 1 1 1 1 1 1 Muscidae sp. C 1 Diptera* 1 1 1 Procladius paludicola 1 1 1 1 1 1 1 1 1 1 Ablabesmyia notabilis 1 Pentaneurini sp. 1 1 Tanytarsus barbitarsis 1 1 Tanytarsus spp. 1 1 1 1 1 1 1 1 1 1 1 Chironomus aff. alternans 1 1 1 1 1 1 1 1 Dicrotendipes 'CA1' Pilbara type 2 1 Dicrotendipes 'CA1' Pilbara type 1 1 Kiefferulus intertinctus 1 1 1 1 1 Polypedilum nubifer 1 1 1 Polypedilum nr. convexum 1 1 1 Cryptochironomus griseidorsum 1

EPHEMEROPTERA Cloeon sp. 1 1 1 1 1 1 1 1 1

HEMIPTERA Mesovelia vittigera 1 1 1 1 Mesoveliidae (juvenile)* 1 Hebrus axillaris 1 1 Microvelia (Pacificovelia) oceanica 1 1 1 Microvelia (Austromicrovelia) peramoena 1 Microvelia (Pacificovelia) lilliput 1 1 Microvelia sp.* 1 1 1 1 Limnogonus (limnogonus) fossarum gilguy 1 1 1 1

51 Gerridae* 1 1 1 Diplonychus eques 1 1 1 1 1 1 Agraptocorixa eurynome 1 Agraptocorixa parvipunctata 1 1 Micronecta gracilis 1 Micronecta quadristrigata 1 1 Micronecta virgata 1 1 1 1 1 1 1 Micronecta sp.* 1 1 1 1 Anisops thienemanni 1 Anisops hackeri 1 1 Anisops nasutus 1 1 1 1 Anisops stali 1 Anisops sp.* 1 1 1 1 1

LEPIDOPTERA Pyralidae 1 1

ODONATA Austroagrion sp. 1 1 1 1 1 1 Ischnura aurora aurora 1 1 1 1 1 1 1 1 1 Xanthagrion erythroneurum 1 1 1 1 1 1 Hemianax papuensis 1 1 1 1 1 1 1 1 Crocothemis nigrifrons 1 Diplacodes bipunctata/trivialis 1 1 1 1 1 1 1 1 1 Orthetrum caledonicum 1 1 1 1 1 1 1 1 1 1 1 Pantala flavescens 1

Tramea stenoloba/loewii 1 1 Libellulidae sp. (juvenile)* 1 1 Hemicordulia tau 1 1 1 1 1

TRICHOPTERA Oecetis sp. 1 1 1 1 Triplectides sp. 1 1 1 1 1 1 1

52