Contributions to Zoology 88 (2019) 452-497 CTOZ brill.com/ctoz
The role of allopatric speciation and ancient origins of Bathynellidae (Crustacea) in the Pilbara (Western Australia): two new genera from the De Grey River catchment
Giulia Perina Centre for Ecosystem Management, School of Science, Edith Cowan University, 270 Joondalup Dv, Joondalup, WA 6027, Australia Western Australian Museum, Locked Bag 49, Welshpool DC, Western Australia 6986, Australia [email protected]
Ana I. Camacho Museo Nacional de Ciencias Naturales (CSIC), Dpto. Biodiversidad y Biología Evolutiva C/ José Gutiérrez Abascal 2, 28006-Madrid, Spain
Joel Huey Centre for Ecosystem Management, School of Science, Edith Cowan University, 270 Joondalup Dv, Joondalup, WA 6027, Australia Western Australian Museum, Locked Bag 49, Welshpool DC, Western Australia 6986, Australia The University of Western Australia, 35 Stirling Hwy, Crawley WA 6009, Australia
Pierre Horwitz Centre for Ecosystem Management, School of Science, Edith Cowan University, 270 Joondalup Dv, Joondalup, WA 6027, Australia
Annette Koenders Centre for Ecosystem Management, School of Science, Edith Cowan University, 270 Joondalup Dv, Joondalup, WA 6027, Australia
Abstract The stygofaunal family of Bathynellidae, is an excellent group to study the processes that shape diversity and distribution, since they have unknown surface or marine relatives, high level of endemism, and limited dispersal abilities. Recent research on Bathynellidae in Western Australia (Pilbara) has uncovered new taxa with unexpected distributions and phylogenetic relationships, but the biogeographical processes that drive their diversification on the continent are still unclear. By exploring the diversity, distribution, and
© Perina et al., 2019 | doi:10.1163/18759866-20191412 This is an open access article distributed under the terms of the cc-by 4.0 License. Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Keywords
ABGD – Anguillanella gen. nov – Bathynellidae – new species – Muccanella gen. nov – PTP – stygofauna
Introduction Aquatic subterranean animals (stygobi- onts) often have larger distributions than ter- Subterranean species distributions are of- restrial subterranean animals (troglobionts) ten restricted, and confined to particular ge- (Barr & Holsinger, 1985; Christman & Culver, ologies, for example isolated karst systems, 2001; Lamoreaux, 2004). This can be due to a limestone, fractured banded iron formations larger suitable continuous environment, such (BIFs), and porous hyporheic systems (Culver as the presence of groundwater in different & Pipan, 2008; Harvey, 2002; Humphreys, 2017; but adjacent geological layers, or to flood- Trontelj et al., 2009). For this reason many sub- ing events that would increase the opportu- terranean taxa are considered short-range en- nity for dispersal (Lamoreaux, 2004). Small demics (SREs; sensu Harvey, 2002). Their small stygobionts are collected not only from karst geographic ranges, and high rate of endemism areas, but also from saturated interstices in make them ideal candidates to explore histori- alluvial/colluvial deposits (Barr & Holsinger, cal climatic and geological events. For example 1985; Holsinger, 1988; Marmonier et al., 1993; ice sheet coverage during the Quaternary gla- Christman & Culver, 2001). In other cases, ciations influenced the distribution of the sub- distributions of stygofaunal species can be re- terranean amphipod species of Stygobromus in stricted to small areas (Eberhard et al., 2009). north America (Holsinger et al., 1997) and Niph- Geographically close aquifers can be isolated argus virei in France (Foulquier et al., 2008). The by impermeable or semi-impermeable mate- occurrence of the cirolanid isopod Antrolana rial such as clay, mudstone or other deposits lira in areas formerly submerged by the ocean, with very low hydraulic conductivity. Ac- confirms the role of marine transgression in cordingly, adjacent groundwater bodies can shaping cirolanid distributions (Holsinger harbour different stygofaunal species. For et al., 1994), and the presence of troglobitic sal- example, calcrete aquifers in ancient palaeo- amanders in Texan caves suggests the ancestor channels of the Yilgarn region, in the arid of this radiation migrated underground to sur- zone of Western Australia, are considered vive climate aridification (Sweet, 1982). “islands under theDownloaded desert” from and Brill.com10/04/2021 host different 08:11:12AM via free access
Figure 1 The Pilbara bioregion with the five major catchments. In red the Goldsworthy study area in the De Grey River catchment, east of Port Hedland town.
partially isolated from the regional alluvial River catchment, we aim to test two hypoth- aquifer (Hickman et al., 1983; Dames & Moore, eses. 1) a single vicariant event (for example a 1992). In the past these perched aquifers were marine regression) isolated the perched aqui- interconnected by higher water table during fers at the same time, and therefore stygofau- the Upper Cretaceous/Early Tertiary (Van na, in different ridges and allopatric specia- de Graaff et al., 1977; BMR Palaeogeographic tion occurred. The phylogeny expected would Group, 1990; González-Álvarez et al., 2016), then show short internodes amongst the taxa and by marine transgressions which occurred identified, and low node support. 2) several in different ages (Artinksian, Oxfordian, Ap- vicariant events (geological events such as tian) (Hickman et al., 1983), when dispersal valley erosion and water level fluctuations) between aquifers, or the origin of a marine fragmented the ancestral population gradu- ancestor into the system, may have been fa- ally in different ridges at different times. We cilitated. By exploring patterns of diversity would then expect multiple ancestors (nodes) (through morphological and molecular tools), at different times, with bathynellids occurring species distribution, and divergence time of on the ridge separated by a deeper valley ge-
Bathynellidae in the north of the De Grey netically more distinctDownloaded from from Brill.com10/04/2021 the others. 08:11:12AM via free access
Material and methods of the area was submerged by marine trans- gressions, for example in the Artinksian, Ox- Study area and groundwater sampling fordian, and Aptian (Hickman et al., 1983). methods Connection and isolation of the perched aqui- The study area (the former Goldsworthy fers probably occurred at different times, but mining area) is located 170 km East of Port with aridification processes started in the Mid- Hedland, in the north of the De Grey River Miocene in the north of Western Australia (Van catchment (fig. 1), one of the main catchments de Graaff et al., 1977; Macphail & Stone, 2004; of the Pilbara bioregion, in the arid zones of Byrne et al., 2008), water tables have dropped WA. It comprises low hills, plateaus alternate steadily, isolating the aquifers in the area. with sandy plains and sporadic claypans. Dif- In 2009 BHP Billiton proposed to mine Cal- ferent ridges of about 260 m Australian Height lawa and Cundaline ridges, therefore sub- Datum are part of the Cleaverville Iron For- terranean fauna surveys were carried out to mation (such as Callawa, Cundaline and Yar- contribute to the development of advice and rie ridges), fractured and weathered Archean- recommendations to the Minister of the Envi- aged formation (Hickman et al., 1983; Williams, ronment on the proposal (Environmental Pro- 2003). Small and large gaps (for example Shay tection Authority, 2007, 2009). A preliminary and Kimberley Gap) between the ridges allow molecular analysis conducted on Bathynel- the passage of many ephemeral creeks that lidae identified two lineages from Cundaline run through the area (Dames & Moore, 1992). and Yarrie ridges respectively (Helix Molecular Aquifers occur in the alluvial deposits, mainly Solution, 2009), but specimens from Callawa in the De Grey River and Eel Creek, and in the ridge were not included in the analysis, there- Archean bedrock that forms the ridges in the fore uncertainties on how many taxa are pres- area. There is evidence of a perched aquifer ent in the area, and their relationships were on Yarrie Ridge confined by granite and mud- unresolved (Subterranean Ecology, unpubl. stone layers that underlie the BIF (see figure data). 3.2 in Dames & Moore, 1992). Rainfall recharg- Given the presence of Bathynellidae in es the perched aquifer through infiltration of different ridges of the Cleaverville Iron For- the permeable BIF on the plateau, but the low mation, the water table fluctuations over geo- hydraulic conductivity of the beneath layers logical time, the drought climate established makes the discharge into the regional water by the Pliocene (Macphail & Stone, 2004) and in the alluvial of the De Grey and Eel Creek the perched aquifers subsequently partially very slow (Dames & Moore, 1992; Williams, isolated from one another and from regional 2003). All ridges of the Cleaverville Forma- groundwater, we believe that this Goldswor- tion abut granitic rocks (Hickman et al., 1983) thy area is an ideal setting to test for allopatric therefore groundwater contained in the BIF speciation in stygofauna. is partially isolated from the regional ground- The bathynellids collected in the Goldswor- water. In geological time frames, water tables thy area during surveys between 2007 and have fluctuated much more than today, inter- 2009 by Subterranean Ecology is the source connecting the perched aquifers, with wetter of the material used for this study. Stygofauna periods of higher water levels, for example in was sampled according to the EPA guidelines the Cretaceous-Early Tertiary (BMR Palaeo- (Environmental Protection Authority, 2003, geographic Group, 1990; González-Álvarez 2007), while specimen preservation follows et al., 2016), and other periods where most Perina et al. (2018).
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Marker COI 16S 16S 18Si 18Sii 18Siii 28S ITS2 TABLE 1 Downloaded from Brill.com10/04/2021 08:11:12AM via free access
DNA methods and data analysis alignment of the five genes considered (Con- Tissue sampling, DNA extraction, PCR ampli- catenate_tree; table 2) were constructed us- fication, and DNA sequencing follow the pro- ing Bayesian and Maximum Likelihood (ML) cedures described in Perina et al. (2018). The method, implemented in RaxML_HPC_Black- mitochondrial Cytochrome oxidase I (COI), Box (Randomized Axelerated Maximum Like- and 16S ribosomal RNA (16S), and the nucle- lihood, including automatic bootstrapping ar 28S, 18S ribosomal RNA, and the Internal stop, which calculates the optimal number of Transcribed Spacer 2 (ITS2) markers were am- replicates to obtain stable support value using plified using the primers listed in table 1. the MRE-based bootstrapping criterion (Pat- We used LIMS (Laboratory Information tengale et al., 2009), and MrBayes on XSEDE Management Software) Biocode plug-in (3.2.6) in the CIPRES Science Gateway online (http://www.mooreabiocode.org) to manage server (Miller et al., 2010), and MrBayes 3.2.5 the molecular laboratory workflows. We im- (Ronquist et al., 2012) respectively. Pilbaranel- ported the raw chromatograms into Geneious la ethelensis was used as outgroup for all single 10.2.4 software (Kearse et al., 2012), and assem- gene trees, but the ITS2 tree, which was rooted bled forward and reverse sequences. Each se- at midpoint. Pilbaranella ethelensis is closely quence was examined by eye and edited. The related to the ingroup taxa, but is a distinct consensus sequences were generated and genus (Perina et al., 2018). Table 2 summarises blasted against GenBank, and sequences the datasets constructed, the best models of representing contaminations were discard- nucleotide substitution for each alignment ed from the analyses. The MAFFT (Multiple selected by the Akaike information criterion Alignment using Fast Fourier Transform) al- in jModeltest 2.1.9 (Posada, 2008), and the gorithm (Katoh et al., 2002) with default pa- analyses performed. rameters was used to build the alignments. We Automatic Barcode Gap Discovery (ABGD) used the online server GBlocks Version 0.91b (Puillandre et al., 2012) and Poisson Tree (Castresana, 2000) to exclude poorly aligned Processes (PTP) (Zhang et al., 2013) species sections of the 16S, ITS2, 18S and 28S align- delimitation methods were implemented, ments using the less strict parameters. Se- using the online websites (http://wwwabi. quences of the mitochondrial COI marker were snv.jussieu.fr/public/abgd/abgdweb.html and translated into amino acid chains to ensure no http://sco.h-its.org/exelixis/web/software/ stop-codons were present. COI fragments pre- PTP/; 13 March 2019) to evaluate hypothetical senting internal stop codons (and therefore species boundaries using phylogenetic trees not coding for the protein) were not consid- and alignments listed in table 2. Kimura 80 ered in the analyses. COI uncorrected pairwise (K80) and Jukes-Cantor (JC69) distance, with distance (P-distance) was calculated within default values, were used in the ABGD analy- and between species haplotypes from dif- sis. Trees had distant outgroups removed to ferent bores (computed through Molecular improve the results, and were analysed to the Evolutionary Genetics Analysis (MEGA) 7.0 PTP method, leaving other settings unvaried. (Kumar et al., 2016) using 1000 bootstraps rep- The molecular phylogeny based on the con- lications and other parameters unvaried) to catenate tree (Concatenate_tree; table 2) was compare with former analyses done on constructed to explore the phylogenetic rela- bathynellids and other groups of stygofauna. tionships among the new species, previously Gene trees based on each gene alignment, defined taxa from the Pilbara, and publicly and a phylogeny based on the concatenated available data from other countries to put the
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
ABGD and PTP in fig. 2) (results ABGD and PTP in fig. 2) (results ABGD and PTP in fig. 2) (results ‒ ‒ ‒ ‒ Analysis ABGD and PTP in fig. 2) (results HKY+G HKY+G GTR+I GTR+G+I HKY+G GTR+G GTR+I Best model selected by by Best model selected (AIC/BIC) jModeltest GTR+G
, , - , Central , Central
from from (align Pilbaranella Pilbaranella Pilbaranella Pilbaranella Pilbaranella Pilbaranella Bathynella Pilbaranella ethelensis Pilbaranella Iberobathynella imuniensis Iberobathynella alignment/tree ofalignment/tree sequences of COI specimens from and one Cundaline ridges Callawa, ofalignment/tree 16S sequences of specimens from and one ridges Yarrie Cundaline, Callawa, ethelensis ofalignment/tree 28S sequences of specimens from and one ridges Yarrie Cundaline, Callawa, ethelensis ofalignment/tree ITS2 sequences of specimens from ridges Yarrie Cundaline, and Callawa, alignment of sequences of COI Callawa, specimens from of representatives Cundaline ridges, and taxa, Gallobathynellinae Qld, SA Hamersley Range, and Bathynellinae, ment trimmed at 610 bp) alignment of 16S sequences of Callawa, specimens from of representatives ridges, Yarrie Cundaline, taxa, and Hamersley Range Central Slovenia alignment of 28S sequences of Callawa, specimens from of representatives ridges, Yarrie Cundaline, taxa Hamersley Range and Central alignment of ITS2 sequences of specimens from ridges Yarrie Cundaline, Callawa, Description Datasets constructed with description, the best model selected by jModeltest, analyses done and number of jModeltest, by with description, the best model selected sequences used constructed Datasets
Dataset COI_alignment/tree 16S_alignment/tree 28S_alignment/tree ITS2_alignment/tree COI_alignment_for tree concatenate 16S_alignment_for tree concatenate 28S_alignment_for tree concatenate ITS2_alignment_for tree concatenate TABLE 2
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
, Central , Central species, species, species, species, species, and species, Pilbaranella Pilbaranella Pilbaranella Pilbaranella Pilbaranella Iberobathynella imuniensis Iberobathynella Fortescuenella serenitatis Fortescuenella serenitatis Fortescuenella serenitatis Fortescuenella alignment of 18S sequences of Callawa, specimens from of representatives Cundaline ridges, and taxa, Gallobathynellinae SA QLD, Hamersley Range, and Bathynellinae, alignment of using the concatenate constructed Tree COI-16S-28S-ITS2-18S genes of alignment with representatives species from COI ridges, Cundaline, Callawa serenitatis Fortescuenella of 16S alignment with representatives species from ridges, Yarrie Cundaline, Callawa, and of 26S alignment with representatives species from ridges, Yarrie Cundaline, Callawa, and of 18S alignment with representatives species from ridges, Yarrie Cundaline, Callawa, and alignment of Concatenate 16S, 28S and 18S genes COI, analysis used in the BEAST Datasets constructed with description, the best model selected by jModeltest, analyses done and number of jModeltest, by with description, the best model selected sequences used ( constructed Datasets
TABLE 2 18S_alignment_for tree concatenate Concatenate_tree BEAST COI_8seq_for 16S_10seq_for BEAST 28S_10seq_for BEAST 18S_10seq_for BEAST Concat_COI-16S-28S- 18S_BEAST_10seq
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Australian fauna into a worldwide context. rates calculated using the highest and lowest Iberobathynella imuniensis (Parabathynelli- rate available, for each marker, retrieved from dae) was used as outgroup. The analysis was the literature from other crustaceans. The 16S partitioned by genes with unlinked models. divergence rates of 0.53% and 1.36% Ma (Still- Between 200,000 and 500,000 Markov Chain man & Reeb, 2001) and the COI divergence Monte Carlo (MCMC) generations were run rates of 1.4% (Knowlton & Weigt, 1998) and for the single gene trees, and 2 × 106 MCMC 2.76% Ma (Wares & Cunningham, 2001) were generations were run for the concatenate tree. used to calculate the ucld.mean and sigma of A burnin fraction of 0.25 was chosen and the the analysis. For the 16S ucld.mean we chose consensus tree was built from the remain- a normal distribution with mean substitution ing trees. The program Tracer 1.6 (Rambaut rate = 0.00473 and sigma = 0.00126 (therefore et al., 2014) was used to assess the convergence 5% Quantile = 0.00266 (corresponding to the of the Bayesian analysis, making sure that the 0.533% divergence rate) and 95% Quantile = Effective Sample Size (ESS) was above 200. 0.0068 (corresponding to the 1.36% diver- The time of divergence of the Goldsworthy gence rate). For the COI ucld.mean we chose taxa was estimated to establish the date of the a normal distribution with mean substitution node that represents the ancestor that colo- rate = 0.0104 and sigma = 0.00205 (therefore nized the perched aquifers, and to establish 5% Quantile = 0.00703 (corresponding to the the origins of Bathynellidae in the area. The 1.4% divergence rate) and 95% Quantile = analysis was conducted using a Bayesian evo- 0.0138 (corresponding to the 2.76% diver- lutionary approach implemented in BEAST gence rate). The ucld.Stdev of both COI and 2.4.8 program (Bouckaert et al., 2014) run in 16S was set as exponential with mean = 1.5. the CIPRES Science Gateway online server 5 × 107 Monte Carlo Markov chains (MCMC) (Miller et al., 2010). The concatenated COI- were run through BEAST 2.4.8 (Bouckaert 16S-28S-18S alignment used in the analysis in- et al., 2014) and convergence was assessed cluded 10 sequences with the most complete using Tracer 1.7 (Rambaut et al., 2018), ensur- dataset. One specimen per species occurring ing the Effective Sample Size (ESS) was above in the study area, representatives of Pilba- 200. Results of the BEAST analysis were sum- ranella species, and Fortescuenella serenitatis marized and annotated using TreeAnnotator as outgroup, a distantly related genus within 2.4.7 (Bouckaert et al., 2014) in a Maximum the same bioregion (Perina et al., 2019). In the Clade Credibility tree, setting a 10% burnin, case of incomplete data, a chimera was as- 0.5 posterior probability limit, and mean node sembled using different specimens belonging heights. The resulting tree was edited in Fig- to the same species. The data were partitioned tree 1.4.2 (http://tree.bio.ed.ac.uk/software/ by gene with unlinked site and clock models, figtree/). linked trees and Yule model. Nucleotide sub- stitution models selected for each gene by the Morphological analysis Akaike information criterion in jModeltest The morphological study follows the methods are listed in table 2. We excluded the propor- described in Perina & Camacho (2016) and Pe- tion of invariant site from the COI, 28S and rina et al. (2018). The material is vouchered at 18S models to simplify the analysis and obtain the Western Australian Museum (see Appen- convergence. A relaxed lognormal clock was dix 2 for voucher numbers). set for all genes. Since no Bathynellacea fossils We used the terminology proposed by Ser- are available to calibrate the molecular clock, ban (1972). The morphological and molecular we utilized 16S and COI mean substitution descriptions are Downloadedbased on from the Brill.com10/04/2021 type series. 08:11:12AM via free access
Abbreviations used in text and figures after are deposited in GenBank (Appendix 2). The Camacho (1986): Th, thoracopod; A.I, anten- ITS2 locus was sequenced for the first time nule; A.II, antenna; Md, mandible, Mx.I, max- for bathynellids as a supplementary locus to illule and Mx.II maxilla. enable more accurate species delineation. The ITS2 primers used for Anguillanella cal- Results lawaensis gen. et sp. nov. were also tested on several specimens of Muccanella cundalinen- The results revealed the presence of two new sis with no success. genera and four new species. The type spe- Species delimitation cies of the two new genera are described in Bayesian single gene trees for COI, 16S, 28S, Appendix 1 (Anguillanella callawaensis gen. and ITS2 together with the species delimi- et sp. nov. and Muccanella cundalinensis tation results for ABGD (with intraspecific gen. et sp. nov.) and for ease of communica- divergence (P) intervals) and PTP are repre- tion, these names are used from this point sented in fig. 2. Specimens from Yarrie ridge forward. failed to sequence for COI, while specimens from Cundaline failed to sequence for ITS2. Molecular results Muccanella cundalinensis was identified by Sixty-five specimens from Callawa, Cundaline morphology and both PTP and ABGD spe- and Yarrie ridges were sequenced and includ- cies delimitation methods applied to COI and ed in the phylogeny. The total number of spec- 28S, while ABGD method using 16S dataset re- imens extracted and successfully amplified turned two groups. for each marker are summarised in table 3. Anguillanella callawaensis was recognised Twenty-one sequences (up to 610 bp) were ob- by morphology and by both methods applied tained for COI. All COI fragments were trans- to 28S and ITS2. However, ABGD of COI align- lated and revealed no stop codons. Forty-five ment split the species into four (0.001 < P < sequences up to 375 bp for the 16S mitochon- 0.0129) and two (0.0215 < P < 0.19) taxa, ABGD drial fragment, fifty-six sequences of about of 16S identified five groups (0.001 < P < 0.0077, 1000 bp for 28S, thirty-three sequences up to with P = prior intraspecific divergence), and 740 bp for ITS2, and forty-three sequences up PTP defined three and two groups respective- to 1780 bp for 18S were obtained. Sequences ly for COI and 16S (fig. 2A-B). Anguillanella TABLE 3 Number of tested and successfully sp. 1 was consistently identified by morpho- amplified specimens per marker, and % logical and molecular data; Anguillanella sp. of success 2 is represented by one specimen only and Marker No No specimens % both methods and all markers identified it as specimens successfully success a distinct species, although a morphological extracted amplified comparison was not possible since the only individual was a juvenile. COI 66 21 31.8 Results of the COI uncorrected P-distance 16S 46 45 97.8 are shown in table 4. P-distance among hap- 18Si 43 43 100.0 lotypes from different bores (within the same 18Sii 43 43 100.0 ridge) ranged between 0 to 16.3%, and within 18Siii 43 37 86.0 same bore holes ranged between 0 to 0.9% in 28S 60 56 93.3 Anguillanella callawaensis gen. et sp. nov. COI ITS2 59 33 55.9 P-distance betweenDownloaded and within from Brill.com10/04/2021 haplotypes 08:11:12AM via free access
Figure 2 Bayesian consensus single gene trees for COI, 16S, 28S and ITS2. Numbers on branches represent Bayesian posterior probabilities followed by maximum likelihood bootstrap percentage. ABGD and PTP results are reported next to the trees. ABGD method: major partitions are showed; PTP: partitions with the highest support for each group are represented.
from the same and different bore holes in slightly different topologies, but both de- Muccanella cundalinensis gen. et sp. nov was fined by three major monophyletic and well 0.1%. Supplementary table S1 provides COI supported groups: Gallobathynellinae and P-distances among all sequences obtained Bathynellinae subfamilies, and the Australian from specimens occurring in Callawa and bathynellids (fig. 3, supplementary fig. S1). The Cundaline ridges. Ten bathynellids from Yar- Australian clade is formed by: two taxa from rie ridge were amplified for COI but resulted South Australia and two from Queensland, in contaminations. The mean pairwise dis- Fortescuenella serenitatis and the other three tance within Anguillanella callawaensis and lineages from the Central Hamersley Range Muccanella cundalinensis is respectively 8.9% (nr Fortescuenella sp., Bathynellidae indet. and 0.1%, while the mean distance between from Mount Florence and SV0138), five taxa the two taxa was 20.1%. of Pilbaranella genus, Anguillanella calla- waensis gen. et sp. nov., Muccanella cundali- Molecular phylogeny nensis gen. et sp. nov., Anguillanella sp. 1 and Maximum likelihood and Bayesian analyses Anguillanella sp. 2. Muccanella cundalinensis, of the concatenated sequences produced Anguillanella callawaensis and Anguillanella Downloaded from Brill.com10/04/2021 08:11:12AM via free access
sp. 1 form monophyletic and well-supported lineages (Posterior Probability (PP) com- CU0285 0.017 0.017 0.017 0.017 0.018 0.018 0.017 0.017 0.001 ‒ prised between 0.96 and 1, and bootstrap (BS) comprised between 95 and 100). The two species collected from Yarrie Ridge form a non-supported clade (PP = 0.72) sister to An- guillanella callawaensis, while in the RaxML 0.017 0.017 0.017 0.017 0.017 0.017 0.017 0.001 (SE = 0.001) 0.001 CU0046 0.017 analysis Anguillanella sp. 1 results sister to An- guillanella callawaensis, but again the node is not supported (BS = 46, see supplementary fig. S1). Anguillanella genus results sister group with Muccanella cundalinensis (PP/BS = 1/100). 0.015 CA0099 0.015 0.015 0.015 0.015 0.015 0.001 (SE = 0.002) 0.003 0.206 0.207 The clades formed by: Gallobathynellinae, Bathynellinae, Fortescuenella serenitatis and nr Fortescuenella sp., Bathynellidae from SA, and Bathynellidae from QLD are well support- ed (BS > 96, PP = 1), but BS and PP are quite low at deeper nodes. 0.015 CA0021 0.015 0.015 0.015 0.015 0.015 0.001 (SE = 0.001) 0.002 0.206 0.207
Molecular clock analysis 0.007 CA0014 0.007 0.006 0.007 0.000 ‒ 0.157 0.158 0.204 0.204 Figure 4 represents the Maximum Clade Cred- ibility (MCC) tree resulting from the molecular dating analysis. Node bars are represented by 0.007 CA0011 0.007 0.006 0.007 ‒ 0.000 0.157 0.158 0.204 0.204 height 95% Higher Posterior Density (HPD). Node ages are indicated above the bars while posterior probability (PP) is below. A chimera
CA0013 0.005 0.005 0.004 ‒ 0.034 0.034 0.162 0.163 0.198 0.199 was assembled for Pilbaranella sp. C using sequences from two specimens (WAMC57501- WAMC60391). The age of the most recent com- mon ancestor (MRCA) between Anguillanella CA0008 0.005 0.005 0.000 0.012 0.033 0.033 0.155 0.156 0.193 0.194 and Muccanella genera is estimated about 64.8 Ma. The mean divergence time of Anguillanella species is 22.6 Ma ago, while Anguillanella cal- CA0160 0.002 0.000 0.014 0.015 0.026 0.026 0.159 0.159 0.193 0.194 lawaensis and Anguillanella sp. 1 result sister species with node age of 15.2 Ma. The MRCA of all Pilbaranella species is dated at 34.2 Ma ago, while the most recent possible speciation event between P. ethelensis and Pilbaranella Estimates of Evolutionary Divergence over Sequence Pairs between haplotypes from the same bore hole. Diagonally in bold: within bore P-distance with P-distance in bold: within bore hole. Diagonally the same bore haplotypes from between Sequence Pairs over of Divergence Estimates Evolutionary possible sp. D dates back to 11.3 Ma. The diver- CA0006 0.009 (SE = 0.005) 0.004 0.018 0.020 0.030 0.030 0.161 0.162 0.196 0.197 COI error standard the diagonal: Above error. standard gence time between Pilbaranella species and
the Goldsworthy taxa is estimated around 96.7 Ma. TABLE 4 CA0006 CA0160 CA0008 CA0013 CA0011 CA0014 CA0021 CA0099 CU0046 CU0285 Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 3 Bayesian consensus tree representing the known Bathynellidae taxa constructed using COI, 16S, 28S, ITS2 and 18S alignments and model partitioning implemented in MrBayes. Numbers on branches represent Bayesian posterior probabilities followed by maximum likelihood bootstrap percentage. Bathynellinae and Gallobathynellinae clades are collapsed for easier interpretation.
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 4 Maximum Clade Credibility Tree inferred using a concatenate COI, 16S, 28S and 18S alignment using BEAST. Node bars are 95% Higher Posterior Density, scale bar is in million years ago (Ma), starting from present 0. Numbers above bars = node age; numbers below bars (bold) = posterior probability of the node.
Discussion by morphological analyses, although ABGD and PTP applied to mitochondrial mark- De Grey River Bathynellidae diversity ers recognise different species distributed An abundance of material previously col- in different bores (fig. 2A-B). We considered lected during several environmental surveys the mitochondrial variability to represent allowed us to examine the morphological and intraspecific population structure (Perina et molecular inter- and intra-specific variabil- al., 2018, 2019), and we used the morphologi- ity, and describe two new genera, Muccanella cal species concept integrated with the above cundalinensis gen. et sp. nov. and Anguillanel- mentioned species delimitation methods. The la callawaensis gen. et sp. nov. Although there tree in fig. 3 (Concatenate_tree; table 2) pres- is a slow discharge of the perched aquifer into ents the phylogenetic relationships among the regional one (Dames & Moore, 1992), the representatives of known genera and lineages results show different lineages on different of Bathynellidae and specimens belonging ridges, which indicates that stygofauna are to the new genera. Each genus and species isolated in each perched aquifer. identified is reciprocally monophyletic sup- The results of the species delimitation porting morphology and species delimitation methods adopted here (fig. 2) are supported methods.
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Anguillanella callawaensis occurs in the recognize different species, the COI uncor- north-eastern part of Callawa Ridge (fig. 5). rected pairwise distance (P-distance) between The population structure highlighted by the specimens occurring in bores CA0021 and mitochondrial loci is comparable to that CA0099 (western side of the ridge) and the found for Pilbaranella ethelensis, with dif- ones collected from bores located in the east- ferent haplotypes in different bores and ern side (CA0006, CA0008, CA0011 CA0013, similar ones within the same bore. Fortes- CA0014 and CA0160) (fig. 5) is quite high cuenella serenitatis, instead, seems to have (between 15.6 and 16.5%, see supplementary a more complex population structure with fig. S2). This “gap” in COI divergences could highly divergent haplotypes (mean COI un- be due to sampling (the south-western part corrected pairwise distance up to 9% (Pe- of the ridge was not sampled, Subterranean rina et al., 2019)) within some bores and the Ecology unpublished data) and hence uncer- same haplotypes occurring in different bores tainties remain regarding the distribution of (see “4 genera networks” in supplementary this species, or the two lineages are diverging fig. S2). Despite subtle morphological differ- (perhaps due to hydrogeological discontinui- ences among specimens of Anguillanella cal- ties), but not long enough to be detected by lawaensis, and ABGD and PTP methods ap- nuclear markers and clear morphological plied to 28S and ITS2, nuclear markers do not differences. We chose to be conservative and
Figure 5 Bathynellidae species distribution in the Goldsworthy area (Callawa, Cundaline, Yarrie ridges).
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Supplementary material Acknowledgements Supplementary material is available online at: This work was supported by: Australian Gov- https://doi.org/10.6084/m9.figshare. ernment’s Australian Biological Resources 8863475
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
References (Crustacea, Syncarida) from Spain. Graellsia, 69, 179–200. doi:10.3989/graellsia.2013.v69.086 Abrams, K.M., Guzik, M.T., Cooper, S.J.B., Hum- Camacho, A.I., Dorda, B.A. & Rey, I. (2013b) Old phreys, W.F., King, R.A., Cho, J.-L. & Austin, A.D. and new taxonomic tools: description of a new (2012) What lies beneath: molecular phyloge- genus and two new species of Bathynellidae netics and ancestral state reconstruction of the from Spain with morphological and molecular ancient subterranean Australian Parabathynel- characters. J. Nat. Hist., 47, 1–28. doi:10.1080/002 lidae (Syncarida, Crustacea). Mol. Phylogenet. 22933.2013.768361 Evol., 64, 1–15. doi:10.1016/j.ympev.2012.03.010 Camacho, A.I., Mas-Peinado, P., Dorda, B.A., Casa- Barr, T.C. & Holsinger, J.R. (1985) Speciation in do, A., Brancelj, A., Knight, L.R.F.D., Hutchins, cave faunas. Annu. Rev. Ecol. Syst., 16, 313–337. B., Bou, C., Perina, G. & Rey, I. (2018) Molecular doi:10.1146/annurev.es.16.110185.001525 tools unveil an underestimated diversity in a BMR Palaeogeographic Group (1990) Australia: stygofauna family: a preliminary world phylog- Evolution of a Continent. Australian Govern- eny and an updated morphology of Bathynel- ment Publishing Service, Canberra. lidae (Crustacea: Bathynellacea). Zool. J. Linn. Bouckaert, R., Heled, J., Kühnert, D., Vaughan, T., Soc.-Lond., 183, 70–96. doi:10.1093/zoolinnean/ Wu, C.-H., Xie, D., Suchard, M.A., Rambaut, A. & zlx063 Drummond, A. (2014) BEAST 2: a software plat- Camacho, A.I., Rey, I., Dorda, B.A., Machordom, A. form for Bayesian evolutionary analysis. PLoS & Valdecasas, A.G. (2002) A note on the system- Biol., 10. doi:org/10.1371/journal.pcbi.1003537 atic position of the Bathynellacea (Crustacea, Bowler, J.M. (1976) Aridity in Australia: age, ori- Malacostraca) using molecular evidence. Con- gins and expression in aeolian landforms trib. Zool., 71, 123–129. and sediments. Earth Sci. Rev., 12, 279–310. Castresana, J. (2000) Selection of conserved blocks doi:10.1016/0012-8252(76)90008-8 from multiple alignments for their use in phy- Brown, L., Finston, T., Humphreys, G., Eberhard, S. logenetic analysis. Mol. Biol. Evol., 17, 540–552. & Pinder, A. (2015) Groundwater oligochaetes doi:10.1093/oxfordjournals.molbev.a026334 show complex genetic patterns of distribution Cho, J.-L. & Humphreys, W.F. (2010) Ten new in the Pilbara region of Western Australia. In- species of the genus Brevisomabathynella vertebr. Syst., 29, 405–420. doi:10.1071/IS14037 Cho, Park and Ranga Reddy, 2006 (Malacos- Byrne, M., Yeates, D.K., Joseph, L., Kearney, traca, Bathynellacea, Parabathynellidae) from M., Bowler, J., Williams, M.A.J., Cooper, S., Western Australia. J. Nat. Hist., 44, 993–1079. Donnellan, S.C., Keogh, J.S., Leys, R., Melville, doi:10.1080/00222930903537066 J., Murphy, D.J., Porch, N. & Wyrwoll, K.H. Cho, J.-L., Park, J.-G. & Humphreys, W.F. (2005) A new (2008) Birth of a biome: insights into the as- genus and six new species of the Parabathynel- sembly and maintenance of the Australian lidae (Bathynellacea, Syncarida) from the arid zone biota. Mol. Ecol., 17, 4398–4417. Kimberley Region, Western Australia. J. Nat. Hist., doi:10.1111/j.1365-294X.2008.03899.x 39, 2225–2255. doi:10.1080/00222930400014148 Camacho, A.I. (1986) A new species of the genus Christman, M.C. & Culver, D.C. (2001) The re- Hexabathynella (Syncarida, Bathynellacea, lationship between cave biodiversity and Parabathynellidae) from Spain. Bijdr. Dierkd., available habitat. J. Biogeogr., 28, 367–380. 56, 123–131. doi:10.1046/j.1365-2699.2001.00549.x Camacho, A.I., Dorda, B.A. & Rey, I. (2013a) Integrat- Coineau, N. & Camacho, A.I. (2013) Superorder ing DNA and morphological taxonomy to de- Syncarida Packard, 1885. In: J.C. von Vaupel scribe a new species of the family Bathynellidae Klein, M. Charmantier-Daures & F.R. Schram
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
(Eds.) The Crustacea – Treatise on Zoology – impact assessment in Western Australia. Gov- Anatomy, Taxonomy, Biology, Volume 4, pp. 357– ernment of Western Australia. 449. Brill, Leiden/Boston. Environmental Protection Authority (2007) Guid- Cooper, S.J.B., Hinze, S., Leys, R., Watts, C.H.S. & ance for the assessment of environmental Humphreys, W.F. (2002) Islands under the des- factors. Sampling methods and survey con- ert: molecular systematics and evolutionary siderations for subterranean fauna in Western origins of stygobitic water beetles (Coleoptera: Australia. Government of Western Australia. Dytiscidae) from central Western Australia. In- Environmental Protection Authority (2009) Cu vertebr. Syst., 16, 589–598. ndaline and Callawa Mining Operations. Report Cooper, S.J.B., Saint, K.M., Taiti, S., Austin, A.D. & and recommendations. Government of West- Humphreys, W.F. (2008) Subterranean archi- ern Australia, Perth. pelago: mitochondrial DNA phylogeography Finston, T.L., Francis, C.J. & Johnson, M.S. (2009) of stygobitic isopods (Oniscidea: Haloniscus) Biogeography of the stygobitic isopod Pygola- from the Yilgarn region of Western Austra- bis (Malacostraca: Tainisopidae) in the Pilbara, lia. Invertebr. Syst., 22, 195–203. doi:10.1071/IS0 Western Australia: evidence for multiple coloni- 7039 sations of the groundwater. Mol. Phylogenet. Evol., Culver, D. & Pipan, T. (2008) Caves as islands. In: R. 52, 448–460. doi:10.1016/j.ympev.2009.03.006 Gillespie (Ed.) Encyclopedia of Islands. Univer- Finston, T.L., Johnson, M.S., Eberhard, S.M., Cock- sity of California Press, Berkeley, CA. ing, J.S., McRae, J.M., Halse, S.A. & Knott, B. Dames & Moore (1992) Goldsworthy Extention (2011) A new genus and two new species of Project – Phase II – Consultative Environmen- groundwater paramelitid amphipods from the tal Review. Prepared for BHP Iron Ore Limited. Pilbara, Western Australia: a combined mo- Delamare Deboutteville, C. & Serban, E. (1973) A lecular and morphological approach. Rec. West. propos du genre Austrobathynella (Bathynella- Aust. Mus., 26, 154–178. doi:10.18195/issn.0312- cea Malacostraca). Livre du cinquantenaire de 3162.26(2).2011.154-178 l’Istitut de spéléologie “Emile Racovitza”. Colloque Finston, T.L., Johnson, M.S., Humphreys, W.F., national spéléologie, Bucarest, 1971, 175–198. Eberhard, S.M. & Halse, S.A. (2007) Cryptic spe- Department of Water (2010) Pilbara regional wa- ciation in two widespread subterranean amphi- ter plan 2010–2030. Government of Western pod genera reflects historical drainage patterns Australia. in an ancient landscape. Mol. Ecol., 16, 355–365. Department of Water (2016) Groundwater as- doi:10.1111/j.1365-294X.2006.03123.x sessment of the north-west Hamersley Range Finston, T.L., Johnson, M.S. & Knott, B. (2008) A (HG62). Perth. new genus and species of stygobitic paramelitid Eberhard, S.M., Halse, S.A., Williams, M., Scan- amphipod from the Pilbara, Western Australia. lon, M.D., Cocking, J.S. & Barron, H.J. (2009). Rec. West. Aust. Mus., 24, 395–410. doi:10.18195/ Exploring the relationship between sampling issn.0312-3162.24(4).2008 efficiency and short range endemism for Fitts, C.R. (2012) Groundwater Science, Second Edi- groundwater fauna in the Pilbara region, West- tion. Elsevier Science, San Diego. ern Australia. Freshwater Biol., 54, 885–901. Foulquier, A., Malard, F., Lefébure, T., Douady, C.J. doi:10.1111/j.1365-2427.2007.01863.x & Gibert, J. (2008) The imprint of Quaternary Environmental Protection Authority (2003) Guid- glaciers on the present-day distribution of the ance for the assessment of environmental fac- obligate groundwater amphipod Niphargus tors. Consideration of subterranean fauna in virei (Niphargidae). J. Biogeogr., 35, 552–564. groundwater and caves during environmental doi:10.1111/j.1365-2699.2007.01795.x
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Giribet, G., Carranza, S., Baguna’, J., Riutort, M. & Island. Paper presented at the Proceedings of Ribera, C. (1996) First Molecular Evidence for the 12th International Congress of Speliology, the existence of a Tardigrada + Arthropoda Switzerland. clade. Mol. Biol. Evol., 13, 76-84. doi:10.1093/ox- Holsinger, J.R., Hubbard, D.A. & Bowman, T.E. fordjournals.molbev.a025573 (1994) Biogeographic and ecological implica- Goater, S.E. (2009) Are Stygofauna Really Protected tions of newly discovered populations of the in Western Australia? PhD thesis. University of stygobiont isopod crustacean Antrolana lira Western Australia. Bowman (Cirolanidae). J. Nat. Hist., 28, 1047– González-Álvarez, I., Salama, W. & Anand, R.R. 1058. doi:10.1080/00222939400770551 (2016) Sea-level changes and buried islands in a Hong, S.J. & Cho, J.-L. (2009) Three new species of complex coastal palaeolandscape in the South Billibathynella from Western Australia (Crusta- of Western Australia: Implications for green- cea, Syncarida, Parabathynellidae). J. Nat. Hist., field mineral exploration. Ore Geol. Rev., 73, 43, 2365–2390. doi:10.1080/00222930903108702 475–499. doi:10.1016/j.oregeorev.2015.10.002 Humphreys, W.F. (2008) Rising from Down Un- Guzik, M.T., Abrams, K.M., Cooper, S.J.B., der: developments in subterranean biodiver- Humphreys, W.F., Cho, J.-L. & Austin, A.D. sity in Australia from a groundwater fauna (2008) Phylogeography of the ancient Para- perspective. Invertebr. Syst., 22, 85. doi:10.1071/ bathynellidae (Crustacea : Bathynellacea) is07016 from the Yilgarn region of Western Austra- Humphreys, W.F. (2017) Australasian subterranean lia. Invertebr. Syst., 22, 205–216. doi:10.1071/ biogeography. In: M.C. Ebach (Ed.) Handbook IS07040 of Australasian Biogeography. CRC Press, Boca Halse, S.A. (2018) Subterranean fauna of the arid Raton, FL. zone. In: H. Lambers (Ed.) On the Ecology of Humphreys, W.F., Watts, C.H.S., Cooper, S.J.B. & Australia’s Arid Zone, pp. 215–241. Springer In- Leijs, R. (2009) Groundwater estuaries of salt ternational Publishing, Cham. lakes: buried pools of endemic biodiversity on Harvey, M.S. (2002) Short-range endemism among the western plateau, Australia. Hydrobiologia, the Australian fauna: some examples from non- 626, 79–95. doi:10.1007/s10750-009-9738-4 marine environments. Invertebr. Syst., 16, 555– Johnson, S.L. & Wright, A.H. (2001) Central Pilbara 570. doi:10.1071/IS02009 groundwater study (Report HG8). Hydrogeo- Helix Molecular Solution (2009) To confirm rela- logical Record Series. Water and Rivers Com- tionships among bathynellids at Cundaline, mission, Perth. Callawa and Yarrie Ridge catchments. Unpub- Karanovic, T. & Cooper, S.J.B. (2011) Molecular lished report. and morphological evidence for short range Hickman, A.H., Chin, R.J. & Gibson, D.L. (1983) endemism in the Kinnecaris solitaria complex Yarrie Western Australia. 1:250 000 Geological (Copepoda: Parastenocarididae), with descrip- Series – Explanatory Notes. Department of Re- tions of seven new species. Zootaxa, 3026, 1–64. sources & Energy. doi:10.11646/zootaxa.3026.1.1 Holsinger, J.R. (1988) Troglobites: the evolution Karanovic, T. & Cooper, S.J.B. (2012) Explosive radi- of cave-dwelling organisms. Am. Sci., 76, 147– ation of the genus Schizopera on a small subter- 153. ranean island in Western Australia (Copepoda: Holsinger, J.R., Carlson, K.R. & Shaw, D.P. (1997) Harpacticoida): unravelling the cases of cryptic Biogeographic significance of recently discov- speciation, size differentiation and multiple in- ered amphipod crustaceans (Stygobromus) in vasions. Invertebr. Syst., 26, 115–192. doi:10.1071/ caves of southeastern Alaska and Vancouver IS11027
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Katoh, K., Misawa, K., Kuma, K. & Miyata, T. (2002) Macphail, M.K. & Stone, M.S. (2004) Age and pal- MAFFT: a novel method for rapid multiple se- aeoenvironmental constraints on the genesis of quence alignment based on fast Fourier trans- the Yandi channel iron deposits, Marillana For- form. Nucleic Acids Res., 30, 3059–3066. mation, Pilbara, northwestern Australia*. Aust. J. Keable, S.J. & Wilson, D.F. (2006) New species of Earth Sci., 51, 497–520. doi:10.1111/j.1400-0952.2004 Pygolabis Wilson 2003 (Isopoda: Tainisopidae, .01071.x Crustacea) from Western Australia. Zootaxa, Marmonier, P., Vervier, P., Gibert, J. & Dole-Olivier, 1116, 1–27. doi:10.5281/zenodo.171623 M.-J. (1993) Biodiversity in ground waters. Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Trends Ecol. Evol., 8, 392–395. Cheung, M., Sturrock, S., Buxton, S., Cooper, Miller, M.A., Pfeiffer, W. & Schwartz, T. (2010) Cre- A., Markowitz, S., Duran, C., Thierer, T., Ash- ating the CIPRES Science Gateway for Inference ton, B., Meintjes, P. & Drummond, A. (2012) of Large Phylogenetic Trees. Paper presented at Geneious Basic: an integrated and extendable the Proceedings of the Gateway Computing En- desktop software platform for the organization vironments Workshop (GCE), New Orleans, LA. and analysis of sequence data. Bioinformat- doi:10.1109/GCE.2010.5676129 ics, 28, 1647–1649. doi:10.1093/bioinformatics/ Nunn, G.B., Theisen, B.F., Christensen, B. & Arc- bts199 tander, P. (1996) Simplicity-correlated size Knott, B. & Halse, S.A. (1999) Pilbarophreatoicus growth of the nuclear 28S ribosomal RNA D3 platyarthricus n.gen., n.sp. (Isopoda: Phreato- expansion segment in the crustacean order icidae: Amphisopodidae) from the Pilbara isopoda. J. Mol. Evol., 42, 211–223. doi:10.1007/ region of Western Australia. Rec. West. Austr. BF02198847 Mus., 51, 33–42. doi:10.3853/j.0067-1975.51.1999 Palumbi, S.R., Martin, A.P., Romano, S., McMillan, .1291 W.O., Stice, L. & Grabowski, G. (1991) The Sim- Knowlton, N. & Weigt, L.A. (1998) New dates and ples Fool’s Guide to PCR. University of Hawaii, new rates for divergence across the Isthmus of Honolulu. Panama. P. Roy. Soc. B-Biol. Sci., 265, 2257–2257. Park, J.-K. & Foighil, D.Ó. (2000) Sphaeriid and Cor- doi:10.1098/rspb.1998.0568 biculid clams represent separate heterodont bi- Kohn, B.P., Gleadow, A.J.W., Brown, R.W., Galla- valve radiations into freshwater environments. gher, K., O’Sullivan, P.B. & Foster, D.A. (2002) Mol. Phylogenet. Evol., 14, 75–88. doi:10.1006/ Shaping the Australian crust over the last 300 mpev.1999.0691 million years: insights from fission track ther- Pattengale, N.D., Alipour, M., Bininda-Emonds, motectonic imaging and denudation studies O.R.P., Moret, B.M.E. & Stamatakis, A. (2009) of key terranes. Aust. J. Earth Sci., 49, 697–717. How many bootstrap replicates are necessary? doi:10.1046/j.1440-0952.2002.00942.x In: S. Batzoglou (Ed.) Research in Computa- Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: tional Molecular Biology: Proceedings 13th An- Molecular Evolutionary Genetics Analysis ver- nual International Conference, RECOMB 2009, sion 7.0 for bigger datasets. Mol. Biol. Evol., 33, Tucson, AZ, USA, May 18–21, 2009, pp. 184–200. 1870–1874. doi:10.1093/molbev/msw054 Springer, Berlin/Heidelberg. Lamoreaux, J. (2004) Stygobites are more wide- Perina, G. & Camacho, A.I. (2016) Permanent slides ranging than troglobites. J. Cave Karst Stud., 66, for morphological studies of small crustaceans: 18–19. Serban’s method and its variation applied on Little, J., Schmidt, D.J., Cook, B.D., Page, T.J. & Bathynellacea (Malacostraca). Crustaceana, 89, Hughes, J.M. (2016) Diversity and phylogeny of 1161–1173. doi:10.1163/15685403-00003576 south-east Queensland Bathynellacea. Aust. J. Perina, G., Camacho, A.I., Huey, J., Horwitz, P. & Ko-
Zool., 64, 36–47. doi:10.1071/ZO16005 enders, A. (2018) DownloadedUnderstanding from Brill.com10/04/2021 subterranean 08:11:12AM via free access
variability: the first genus of Bathynellidae Lit. Mainz, Math-Nat., Mikrofauna Meeres- (Bathynellacea, Crustacea) from Western Aus- bodens, 24, 1–192. tralia described through a morphological and Schminke, H.K. (1974) Mesozoic intercontinental multigene approach. Invertebr. Syst., 32, 423– relationships as evidenced by bathynellid Crus- 447. doi:10.1071/IS17004 tacea (Syncarida: Malacostraca). Syst. Zool., 23, Perina, G., Camacho, A.I., Huey, J., Horwitz, P. & 157–164. Koenders, A. (2019) New Bathynellidae (Crus- Sedghi, M. (2016) On the hydraulics of Extensive tacea) taxa and their relationships in the For- perched aquifers. Paper presented at the 2nd tescue catchment aquifers of the Pilbara region, National Conference of Water Crisis in Iran and Western Australia. Syst. Biodivers., 17, 148–164. the Middle East. doi:10.1080/14772000.2018.1559892 Serban, E. (1972) Bathynella (Podophallocarida, Pinder, A. (2008) Phreodrilidae (Clitellata: Annel- Bathynellacea). Trav. Instit. Spéol. “É. Racovitza”, ida) in north-western Australia with descrip- 11, 11–225. tions of two new species. Rec. West. Austr. Mus., Simon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, 24, 459–468. doi:10.18195/issn.0312-3162.24(4). H. & Flook, P. (1994) Evolution, weighting, and 2008.459-468 phylogenetic utility of mitochondrial gene- Posada, D. (2008) jModelTest: phylogenetic mod- sequences and a compilation of conserved el averaging. Mol. Biol. Evol., 25, 1253–1256. polymerase chain-reaction primers. Ann. Ento- doi:10.1093/molbev/msn083 mol. Soc. Am., 87, 651–701. doi:10.1093/aesa/87.6. Puillandre, N., Lambert, A., Brouillet, S. & Achaz, 651 G. (2012) ABGD, Automatic Barcode Gap Dis- Smith, G.B., Eberhard, S.M., Perina, G. & Fin- covery for primary species delimitation. Mol. ston, T.L. (2012) New species of short range Ecol., 21, 1864–1877. doi:10.1111/j.1365-294X.2011. endemic troglobitic silverfish (Zygentoma: 05239.x Nicoletiidae) from subterranean habitats in Rambaut, A., Drummond, A.J., Xie, D., Baele, G. & Western Australia’s semi-arid Pilbara region. Suchard, M.A. (2018) Posterior summarization Rec. West. Austr. Mus., 27, 101–116. doi:10.18195/ in Bayesian phylogenetics using tracer 1.7. Syst. issn.0312-3162.27(2).2012.101-116 Biol., 67, 901–904. doi:10.1093/sysbio/syy032 Stillman, J. & Reeb, C. (2001) Molecular phylog- Rambaut, A., Suchard, M.A., Xie, D. & Drummond, eny of Eastern Pacific porcelain crabs, Genera A.J. (2014) Tracer v1.6. Available online: http:// Petrolisthes and Pachycheles, based on the beast.bio.ed.ac.uk/Tracer. mtDNA 16S rDNA sequence: phylogeographic Rix, M.G. (2009) Taxonomy and Systematics of the and systematic implications. Mol. Phylogenet. Australian Micropholcommatidae (Arachnida: Evol., 19, 236–245. doi:10.1006/mpev.2001.0924 Araneae). PhD thesis. The University of West- Sweet, S.S. (1982) A distributional analysis of Epige- ern Australia, Perth. an populations of Eurycea neotenes in Central Ronquist, F., Teslenko, M., van der Mark, P., Ayres, Texas, with comments on the origin of troglo- D.L., Darling, A., Höhna, S., Larget, B., Liu, bitic populations. Herpetologica, 38, 430–444. L., Suchard, M.A. & Huelsenbeck, J.P. (2012) Trontelj, P., Douady, C.J., Fišer, C., Gibert, J., MrBayes 3.2: Efficient Bayesian phylogenetic Gorički, Š., Lefébure, T., Sket, B. & Zakšek, V. inference and model choice across a large (2009) A molecular test for cryptic diversity model space. Syst. Biol., 61, 539–542. doi:10.1093/ in ground water: how large are the ranges of sysbio/sys029 macro-stygobionts? Freshwater Biol., 54, 727– Schminke, H.K. (1973) Evolution, System und Verb- 744. doi:10.1111/j.1365-2427.2007.01877.x reitungsgeschichte der Familie Parabathynelli- Van de Graaff, W.J.E., Crowe, R.W.A., Bunting, J.A.
dae (Bathynellacea, Malacostraca). Akad. Wiss. & Jackson, M.J.Downloaded (1977) fromRelict Brill.com10/04/2021 early Cainozoic 08:11:12AM via free access
drainages in arid Western Australia. Zeitschrift Pilbara, Western Australia. Zootaxa, 245, 1–20. für Geomorphologie, 21, 379–400. doi:10.11646/zootaxa.245.1.1 Wares, J.P. & Cunningham, C.W. (2001) Phylo- WorleyParsons (2012) Pilbara Iron Ore Project – geography and historical ecology of the north Groundwater Impact Assessment Report. Re- atlantic intertidal. Evolution, 55, 2455–2469. port prepared for Flinders Mine Limited, Perth. doi:10.1111/j.0014-3820.2001.tb00760.x Zhang, J., Kapli, P., Pavlidis, P. & Stamatakis, A. Whiting, M.F., Carpenter, J.M. & Wheeler, Q.D. (2013) A general species delimitation method (1997) The Stresiptera problem: phylogeny of with applications to phylogenetic placements. the holometabolous insect orders inferred from Bioinformatics, 29, 2869–2876. doi:10.1093/bioin 18S and 28S ribosomal DNA sequences and mor- formatics/btt499 phology. Syst. Biol., 46, 1–68. doi:10.2307/2413635 Williams, I.R. (2003) Yarrie, W.A. (3rd Edition): RECEIVED: 30 APRIL 2019 | REVISED AND ACCEPTED: Western Australia Geological Survey, 1:250,000, 11 JULY 2019 Geological Series Explanatory Notes, pp. 84. EDITOR: R. VONK Wilson, G.D.F. (2003) A new genus of Tainisopi- dae fam. nov. (Crustacea: Isopoda) from the
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
appendix 1 to five small denticles. Thoracopods I to VII with epipod and endopod four-segmented. Thoracopod Systematic account VIII male rectangular and compact, with only one well developed lobe (outer lobe) on penial region Thirty-nine specimens of Anguillanella calla- (latero external part) and two projections (fron- waensis gen. et sp. nov. were used for DNA extrac- tal and posterior); basipod in vertical position tion and are part of the type series. The extractions with large distal crest; endopod small and exopod were obtained from: 18 whole specimens, and 21 large, perpendicular to basipod and curved out- selected body parts (few body segments where ward, with simplified morphology and two distal no morphological characters are present and/or a setae present. Female thoracopod VIII simplified: piece of the specimen’s upper part of pereionites coxopod, without setae; epipod very long; basipod and pleonites). Sixteen specimens of Muccanella large; one-segmented small ramus (endopod or cundalinensis gen. et sp. nov. have been used for exopod). Uropod: sympod with four inhomono- DNA extraction and are part of the type series. The mous spines and endopod with two spines, one extractions were obtained from eight whole speci- special seta (seta “X”) and three more setae (one mens and eight selected body parts. Selected body very short). Furcal rami with five spines. parts of nine individuals of Anguillanella sp. 1 gen. nov., and one whole specimen of Anguillanella sp. Type species: Anguillanella callawaensis gen. nov., 2 have been sequenced successfully and used in sp. nov the molecular study. Anguillanella callawaensis Perina and Family Bathynellidae Grobben, 1905 Camacho sp. nov. (figs. 6,7,8–9) The Family Bathynellidae comprises three subfam- Zoobank: urn:lsid:zoobank.org:act:2399400E-4AA2 ilies: Bathynellinae Grobben, 1905, Gallobathynel- -4697-80D6-0925B1DAD43E linae Serban, Coineau & Delamare Deboutteville, 1971 and Austrobathynellinae Delamare Deboutte- Type locality. Bore CA0008, Callawa Ridge, De Grey ville & Serban, 1973. River Catchment, Pilbara, Western Australia (see Appendix 3 for borehole coordinates). Anguillanella Perina and Camacho gen. nov. Generic diagnosis Material examined Zoobank: urn:lsid:zoobank.org:act:17F7F477-6B59- Holotype. WAMC57418, male, permanent slide, 458C-9A43-E3496A17B317 bore CA0008, 24 July 2009, Bell (stygo net haul). Allotype. WAMC57370, female, permanent slide, Antennula seven-segmented. Antenna seven-seg- bore CA0157, 10 July 2009, Bell (stygo net haul). mented, third endopodial segment very small. Pa- Paratypes. WAMC57258, female, permanent slide, ragnaths with distal strong claws. Mandibular palp Western Australia, Pilbara, bore CA0006, 13 June three segmented with two long and strong equal 2009, Bell & Ridley (stygo net haul); WAMC57260, barbed claws, and without sexual dimorphism. female, 100% ethanol, Western Australia, Pilbara, Pars incisiva with two teeth; processus incisivus ac- bore CA0006, 13 June 2009, Bell & Ridley (stygo net cessorius with one tooth and one seta-like tooth; haul); WAMC57262, female, 100% ethanol, Western pars molaris with two dentate structures, paral- Australia, Pilbara, bore CA0006, 13 June 2009, Bell lel to main axis of teeth: first structure (closest & Ridley (stygo net haul); WAMC57263, sex not to processus incisivus accessorius) with four teeth, recorded, whole specimen for DNA, Western Aus- and second structure formed by a bulb with three tralia, Pilbara, bore CA0006, 26 April 2008, Bell
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 6 Anguillanella callawaensis gen. et sp. nov., male holotype (A–E, G); female allotype (F, H, K, L); male paratype (I, J). (A) Antennula (dorsal view); (B) antenna (dorsal view); (C) max Maxilla; (D) maxillula; (E) mandibular palp male holotype; (F) palp female allotype; (G) mandible male holotype; (H) mandi- ble female allotype; (I) paragnath male WAMC57657 (J) labrum male WAMC57423 (ventral view); (K) Paragnath female allotype; (L) labrum female allotype (dorsal view). ScaleDownloaded bar in from mm. Brill.com10/04/2021 08:11:12AM via free access
Figure 7 Anguillanella callawaensis gen. et sp. nov., male holotype. (A) Thoracopod I; (B) thoracopod II; (C) thoracopod III; (D) thoracopod IV; (E) thoracopod V; (F) Thoracopod VI; (G) thoracopod VII. Scale bar in mm.
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 8 Anguillanella callawaensis gen. et sp. nov., (A–D, F, G, H) male holotype. (A, B) thoracopod VIII (pos- terior view); (C, D) thoracopod VIII (frontal view); (E) thoracopod VIII female allotype (frontal view); (F) first pleopod; (G) furcal rami and dorsal seta (dorsal view); (H) uropod (latero-internal view). Scale bar in mm. Abbreviations: O. lb, outer lobe; Bsp, basipod; Endp, endopod; Exp, exopod; P.pr, posterior projection; Fr.pr, frontal projection.
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Pilbara, bore CA0012, 31 May 2009, Bell & Bar- net (stygo net haul); WAMC57281, female, whole specimen for DNA, Western Australia, Pilbara, bore CA0012, 31 May 2009, Bell & Barnet (stygo net haul); WAMC57283, female, whole specimen for DNA, Western Australia, Pilbara, bore CA0013, 09 July 2009, Bell (stygo net haul); WAMC57285, male, permanent slide, Western Australia, Pilbara, bore CA0014, 09 July 2009, Bell (stygo net haul); WAMC57286, female, whole specimen for DNA, Western Australia, Pilbara, bore CA0014, 09 July 2009, Bell (stygo net haul);WAMC57288, female, Figure 9 Anguillanella callawaensis furca variability (WAMC57370 photo). permanent slide, Western Australia, Pilbara, bore CA0021, 31 May 2009, Bell & Barnet (stygo net haul); & Eberhard (stygo net haul); WAMC57264, sex WAMC57289, female, whole specimen for DNA, not recorded, whole specimen for DNA, Western Western Australia, Pilbara, bore CA0021, 31 May Australia, Pilbara, bore CA0006, 26 April 2008, 2009, Bell & Barnet (stygo net haul); WAMC57290, Bell & Eberhard (stygo net haul); WAMC57266, female, whole specimen for DNA, Western Aus- male, permanent slide, Western Australia, Pil- tralia, Pilbara, bore CA0021, 31 May 2009, Bell & bara, bore CA0008, 09 July 2009, Bell (stygo net Barnet (stygo net haul); WAMC57371, female, 100% haul); WAMC57267, male, permanent slide, West- ethanol, Western Australia, Pilbara, bore CA0157, ern Australia, Pilbara, bore CA0008, 09 July 2009, 10 July 2009, Bell (stygo net haul); WAMC57373, Bell (stygo net haul); WAMC57269, female, 100% female, 100% ethanol, Western Australia, Pilbara, ethanol, Western Australia, Pilbara, bore CA0008, bore CA0160, 09 July 2009, Bell (stygo net haul); 22 July 2009, Bell (stygo net haul); WAMC57270, WAMC57374, juvenile, whole specimen for DNA, female, 100% ethanol, Western Australia, Pil- Western Australia, Pilbara, bore CA0160, 09 July bara, bore CA0008, 22 July 2009, Bell (stygo net 2009, Bell (stygo net haul); WAMC57375, juvenile, haul); WAMC57271, female, whole specimen for whole specimen for DNA, Western Australia, Pil- DNA, Western Australia, Pilbara, bore CA0008, bara, bore CA0160, 09 July 2009, Bell (stygo net 22 July 2009, Bell (stygo net haul); WAMC57273, haul); WAMC57377, female, 100% ethanol, West- female, permanent slide, Western Australia, Pil- ern Australia, Pilbara, bore CA0099, 13 June 2009, bara, bore CA0011, 23 July 2009, Bell (stygo net Bell & Ridley (stygo net haul); WAMC57378, fe- haul); WAMC57274, sex not recorded, whole male, 100% ethanol, Western Australia, Pilbara, specimen for DNA, Western Australia, Pilbara, bore CA0099, 13 June 2009, Bell & Ridley (stygo net bore CA0011, 23 July 2009, Bell (stygo net haul); haul); WAMC57379, male, permanent slide West- WAMC57275, male, permanent slide, Western ern Australia, Pilbara, bore CA0099, 13 June 2009, Australia, Pilbara, bore CA0011, 23 July 2009, Bell Bell & Ridley (stygo net haul); WAMC57380, fe- (stygo net haul); WAMC57277, female, 100% male, permanent slide Western Australia, Pilbara, ethanol, Western Australia, Pilbara, bore CA0011, bore CA0099, 13 June 2009, Bell & Ridley (stygo 12 June 2009, Bell & Ridley (stygo net haul); net haul); WAMC57414, female, permanent slide WAMC57278, male, whole specimen for DNA, Western Australia, Pilbara, bore CA0008 09 July Western Australia, Pilbara, bore CA0011, 12 June 2009, Bell (stygo net haul); WAMC57415, female, 2009, Bell & Ridley (stygo net haul); WAMC57280, permanent slide Western Australia, Pilbara, female, permanent slide, Western Australia, bore CA0008 09 July 2009, Bell (stygo net haul);
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
WAMC57416, sex not identifiable, permanent Abbreviations used: Th, thoracopod; A.I, anten slide Western Australia, Pilbara, bore CA0008 09 nule; A.II, antenna; Md, mandible, Mx.I, maxillule July 2009, Bell (stygo net haul); WAMC57417, fe- and Mx.II maxilla. male, permanent slide Western Australia, Pilbara, bore CA0008 09 July 2009, Bell (stygo net haul); Description (based on adults) WAMC57419, female, permanent slide Western Body. Total length of holotype and allotype Australia, Pilbara, bore CA0008 09 July 2009, Bell 1 mm. Total length of males 0.7–1 mm, and females (stygo net haul); WAMC57420, male, permanent 0.8–1. mm. Body elongated; almost cylindrical, slide Western Australia, Pilbara, bore CA0008 09 nearly eight times as long as wide; segments slight- July 2009, Bell (stygo net haul); WAMC57421, fe- ly widening towards posterior end. Head slightly male, permanent slide Western Australia, Pilbara, longer than wide. Pleotelson with one plumose bore CA0008 09 July 2009, Bell (stygo net haul); dorsal seta on either side. All drawings are of the WAMC57422, female, permanent slide Western holotype and allotype except: paragnath in fig. 6I, Australia, Pilbara, bore CA0008 09 July 2009, Bell and labrum in fig. 6J. (stygo net haul); WAMC57423, male, permanent Antennule (fig. 6A). Seven-segmented; length of slide Western Australia, Pilbara, bore CA0008 09 first three segments slightly longer than the last July 2009, Bell (stygo net haul); WAMC57424, fe- four; first segment is the longest, then sixth and male, permanent slide Western Australia, Pilbara, seventh, as well as second and third, of similar bore CA0008 09 July 2009, Bell (stygo net haul); length; fourth small and fifth very small; inner fla- WAMC57425, male, permanent slide Western Aus- gellum small and trapezoidal; setation as in fig. 6A; tralia, Pilbara, bore CA0008 09 July 2009, Bell (sty- one and two aesthetascs on segments sixth and go net haul); WAMC57426, juvenile, permanent seventh respectively. slide Western Australia, Pilbara, bore CA0008 09 Antenna (fig. 6B). Seven-segmented; as long as the July 2009, Bell (stygo net haul); WAMC57657, male, first six segments of A.I; first four segments almost permanent slide, Western Australia, Pilbara, bore as long as the last three ones; fifth (third of endo- CA0124, 14 June 2009, Bell & Ridley (stygo net haul); pod) very small, without setae; terminal segment WAMC59191, female, whole specimen for DNA, is the longest; first, fourth and sixth segments simi- Western Australia, Pilbara, bore CA0124, 14 June lar in length and about 0.8 times as long as seventh 2009, Bell & Ridley (stygo net haul); WAMC59192, segment; setal formula: 0/1+exp/2+0/1+0/0/2+0/5; sex not recorded, whole specimen for DNA, West- exopod about twice as long as second seg- ern Australia, Pilbara, bore CA0124, 14 June 2009, ment, with two terminal setae, one smooth and Bell & Ridley (stygo net haul); WAMC59193, fe- one bifurcated sensory seta; ventromedial seta male, whole specimen for DNA, Western Australia, absent. Pilbara, bore CA0124, 14 June 2009, Bell & Ridley Labrum (fig. 6J, L). Almost square, with smooth (stygo net haul); WAMC59194, male, permanent free edge, and with two small lobes, more or less slide, Western Australia, Pilbara, bore CA0057, developed, on the distal part. 23 July 2009, Bell (stygo net haul); WAMC59195, Paragnath (fig. 6I, K). Almost rectangular, strong male, permanent slide, Western Australia, Pilbara, long claw on distal part with thick setation, distal bore CA0057, 23 July 2009, Bell (stygo net haul); ventral section slightly dilated. WAMC59196, female, whole specimen for DNA, Mandible (fig. 6E–H). Palp with three segments, Western Australia, Pilbara, bore CA0057, 23 July terminal segment with two long and strong equal 2009, Bell (stygo net haul); WAMC59197, female, barbed claws, more or less cylindrical without whole specimen for DNA, Western Australia, Pilba- expansions (fig. 6E, F). Masticatory part (fig. 6G, ra, bore CA0057, 23 July 2009, Bell (stygo net haul); H): pars incisiva (incisor process) with two teeth;
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 10 Muccanella cundalinensis gen. et sp. nov., male holotype (A, B, D, F, H, I); female allotype (E, G); male paratype (C). (A) Antennula (dorsal view); (B) antenna (dorsal view); (C) Paragnath male WAMC57343; (D) labrum; (E) labrum female WAMC57341; (F) palp and mandible male holotype; (G) palp and mandible female allotype; (H) maxillule; (I) maxilla. Scale bar in mm.
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Figure 11 Muccanella cundalinensis gen. et sp. nov., male holotype. (A) Thoracopod I; (B) thoracopod II; (C) thora- copod III; (D) thoracopod IV; (E) thoracopod V; (F) Thoracopod VI; (G) Downloadedthoracopod from VII. Brill.com10/04/2021 Scale bar in mm. 08:11:12AM via free access
Figure 12 Muccanella cundalinensis gen. et sp. nov., (A–D, F–H) male holotype; female allotype (E). (A, B) thora- copod VIII (posterior view); (C, D) thoracopod VIII (frontal view); (E) thoracopod VIII female allotype (frontal view); (F) first pleopod; (G) furcal ramus and dorsal seta (dorsal view); (H) uropod (dorsal view). Scale bar in mm. Abbreviations: O. lb, outer lobe; Bsp, basipod;Downloaded Endp, endopod; from Brill.com10/04/2021 Exp, exopod; 08:11:12AM P.pr, posterior projection; Fr.pr, frontal projection. via free access
2009, Bell & Barnet; WAMC57365, female, whole slightly shorter, fifth very small; inner flagel- specimen for DNA, bore CU0058, 29 May 2009, lum small and longer than wide; setation as in Bell & Barnet; WAMC57366, juvenile, whole fig. 10A; two aesthetascs on both sixth and seventh specimen for DNA, bore CU0058, 29 May 2009, segments. Bell & Barnet; WAMC57368, female, permanent Antenna (fig. 10B). Seven-segmented; nearly as slide, bore CU0285, 30 May 2009, Bell & Barnet; long as A.I; first four segments almost as long as WAMC59198, male, permanent slide, bore CU0058, the last three combined; fifth (third of endopod) 10 June 2009, Bell & Ridley; WAMC59199, sex not very small, without setae; terminal segment is the recorded, permanent slide, bore CU0058, 10 June longest, fourth and sixth segments similar in length 2009, Bell & Ridley; WAMC59205, female, perma- and about three-fourth the length of seventh seg- nent slide, bore CU0058, 29 May 2009, Bell & Barnet; ment; setal formula: 0/1+exp/2+0/1+0/0/2+0/5; WAMC59206, male, permanent slide, bore CU0064, exopod 0.8 times the length of the first segment, 21 July 2009, Bell; WAMC59207, male, permanent with two terminal setae, one smooth and one bi- slide, bore CU0064, 21 July 2009, Bell; WAMC59208, furcated sensory seta; ventromedial seta absent. male, permanent slide, bore CU0064, 21 July 2009, Labrum (fig. 10D, E). Almost square, with smooth free Bell; WAMC59209, male, permanent slide, bore edge, and with two small lobes on the distal part. CU0046, 29 May 2009, Bell & Barnet; WAMC59210, Paragnath (fig. 10C). Almost rectangular, strong female, permanent slide, bore CU0046, 29 May long claw on distal part with thick setation. 2009, Bell & Barnet; WAMC59211, male, permanent Mandible (fig. 10F, G). Palp with three segments, slide, bore CU0046, 29 May 2009, Bell & Barnet; terminal segment with two long and strong barbed WAMC59212, male, permanent slide, bore CU0046, claws of similar length, more or less cylindrical 29 May 2009, Bell & Barnet; WAMC59213, female, without expansions. Masticatory part: pars inci- permanent slide, bore CU0046, 29 May 2009, Bell & siva with two teeth; processus incisivus accessorius Barnet; WAMC59214, male, permanent slide, bore with one tooth and one seta-like tooth; pars mo- CU0046, 29 May 2009, Bell & Barnet; WAMC59215, laris with three teeth close to processus incisivus male, permanent slide, bore CU0046, 29 May 2009, accessorius and one distal strong tooth with one Bell & Barnet; denticle on each side. Maxillule (fig. 10H). Proximal endite with four se- Abbreviations used: Th, thoracopod; A.I, anten tae; distal endite with six teeth, four with denticles nule; A.II, antenna; Md, mandible, Mx.I, maxillule and two setae-like, and three plumose setae in and Mx.II maxilla. outer margin of endite. Maxilla (fig. 10I). Four-segmented; setal formula 7, Description (based on adults) 3, 7, 5. Body. Total length of holotype about 0.95 mm and Thoracopods I-VII (fig. 11A–G). Epipod present on allotype 0.94 mm. Total length of males 0.82–1.08 Th I to VII. Th I coxa with a long and strong plu- mm, of females 0.92–1.07 mm. Body almost cylin- mose seta and an external bundle of thin hair; drical, nearly six times as long as wide; segments basipod with two smooth setae. One-segmented slightly widening towards posterior end. Head as exopod, shorter than endopod on all thoracopods; long as wide. Pleotelson with one plumose dorsal exopod reaches about the middle of the third seg- seta on either side. All drawings are of the holo- ment of endopod in all thoracopods; it bears four type and allotype except for fig. 10C, E. barbed setae on Th I, two terminal, one dorsal and Antennule (fig. 10A). Seven-segmented; length of one ventral; and five barbed setae on Th II to VII. first three segments as long as the last three; first Exopod of thoracopos I to VI with tuft of setules and last segments are the longest, then sixth, on ventral margin (on dorsal margin too on second and third of similar length and fourth thoracopod I). EndopodDownloaded with fromfour Brill.com10/04/2021 segments in all08:11:12AM via free access
Figure 13 Male and female thoracopods VIII of the four genera described for WA. (A, B) male ThVIII of Pil- baranella ethelensis; (C, D) male ThVIII of Fortescuenella serenitatis; (E, F) male ThVIII of Anguillanella callawaensis; (G, H) male ThVIII of Muccanella cundalinensis; (I) female ThVIII of Pilbaranella ethelen- sis; (J) female ThVIII of Fortescuenella serenitatis; (K) female ThVIII of Anguillanella callawaensis; (L) female ThVIII of Muccanella cundalinensis. Scale bar in mm
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
WA (fig. 13) but all have endopod and exopod A. callawaensis. Differences could represent intra- reduced, and the latter curved backwards like specific variation or perhaps the population from Austrobathynellinae, while Bathynellinae and these bores is diverging, but not long enough to Gallobathynellinae have unfolded and more de- show clear morphological differences. veloped rami. In Pilbaranella the male thoracopod Bathynella primaustraliensis (Schminke, 1973), VIII is simplified and reduced; in Fortescuenella the only bathynellid species described for Australia it is more complex with a frontal and posterior before 2018 and collected from the Hughes creek in projection and a well-developed outer lobe; in Victoria, differs from the new genera in mandible and Muccanella and Anguillanella we have the same female thoracopod VIII. A comparison of the male structures present in Fortescuenella (frontal- thoracopod VIII (penis) is not possible since the posterior projection and outer lobe), but in Muc- description is based on one female (Schminke, canella they are reduced while they are well- 1973), but we justify the erection of two new genera developed in Anguillanella, and both taxa have ba- based on molecular evidence and that bathynellid sipod with crest. Setation on endopod and exopod species and genera of the Pilbara bioregion seem is the same in all four genera (one short seta on to be restricted to well-defined parts of the catch- endopod and two setae on exopod). ments where they occur (Perina et al., 2018, 2019). Female thoracopod VIII (fig. 13) is uniramus As Pilbaranella and Fortescuenella, we exclude like in Austrobathynella patagonica Delamare the new genera from Bathynellinae and Gallo- Deboutteville & Roland, 1963; it is very simple in bathynellinae subfamilies based on: number of teeth Pilbraranella with most of the segments fused; on pars molaris and structure of male and female in Fortescuenella and Anguillanella it is formed thoracopod VIII, supporting the molecular data. by: coxopod, basipod and one-segmented ramus, The new genera exhibit some common char- while in Muccanella the ramus is two-segmented. acters with members of the Austrobathynellinae All genera so far described for WA present epipod, subfamily, but they are quite distinguished from contrary to Austrobathynella patagonica. members of Bathynellinae and Gallobathynellinae The pleopod I differs in all taxa by the number of (table 5). With Austrobathynella Delamare Debout- setae, which are all quite elongated in Pilbraranella, teville, 1960, Transvaalthynella Serban & Coineau, Anguillanella and Muccanella, while Fortescuenella 1975 and Transkeithynella Serban & Coineau, 1975, presents one very short apical seta. Another charac- genera belonging to Austrobathynellinae, Anguil- ter that distinguishes Fortescuenella is the absence lanella and Muccanella present: many teeth on pars of the “special seta X” (Delamare Deboutteville & molaris of the mandible; female thoracopod VIII Serban, 1973), which is present in the other three reduced to one ramus; male thoracopod VIII with genera. All four taxa have four spines on sympod reduced endopod and exopod; and third segment (but different length), two on endopod of uropod of antennule’s endopod very small (short). The and five on furca with distinct ration length. paucity of data (only three genera are described Specimens from bore YP1063 show affinity to based solely on morphological data) prevents us Anguillanella callawaensis, but also distinctive from corroborating the affinity of the new genera morphological characters. The only specimen Anguillanella and Muccanella to Austrobathynel- sequenced from bore YRP22 on Yarrie Ridge was linae, nevertheless they are more dissimilar from a juvenile wholly used for DNA extraction, so no Bathynellinae and Gallobathynellinae and there- morphological characters are available, but the fore we exclude them. Further morphological and molecular data place it within the “Anguillanella molecular data are needed to outline Bathynelli- group” therefore we included it in this genus. dae subfamilies, resolve taxonomic uncertainties, Specimens from bores CA0021 and CA0099 have and systematically organise the taxa discovered in slightly different morphology from the rest of different continents.Downloaded from Brill.com10/04/2021 08:11:12AM via free access
-
number number of AI: segments AII: of segments endopod seg ment 3 me exopod: dial seta Mandible: palp pars molaris (number of teeth) sexual dimorphism VII: Th I-Th endopod Th VIII female: rami (endop-exop) seta coxal epipod TABLE 5
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
Downloaded from Brill.com10/04/2021 08:11:12AM via free access
appendix 3
Bore holes coordinates
Bore Latitude Longitude CA0006 20°38’40.02”S 120°18’22.57”E CA0008 20°38’33”S 120°18’2”E CA0011 20°38’57”S 120°18’17”E CA0012 20°39’6”S 120°18’23”E CA0013 20° 39’ 12.96”S 120° 18’ 14.47”E CA0014 20° 38’ 51.10”S 120° 18’ 15.33”E CA0021 20° 38’ 41.92”S 120° 17’ 13.23”E CA0057 20°39’37”S 120°18’10”E CA0099 20°38’40”S 120°17’32”E CA0124 20°38’54”S 120°17’57”E CA0157 20°39’25”S 120°17’58”E CA0160 20°38’23”S 120°18’40”E CU0046 20°32’36.46”S 120°09’35.42”E CU0058 20°32’21.8”S 120°09’12.3”E CU0064 20°32’20.61”S 120°09’13.5”E CU0285 20°32’49.38”S 120°10’00.98”E YP1063 20°36’11.74”S 120°20’23.46”E YRP22 20°36’32.22”S 120°18’26.42”E
Downloaded from Brill.com10/04/2021 08:11:12AM via free access