Patterns of population genetic variation in sympatric chiltoniid amphipods within a calcrete aquifer reveal a dynamic subterranean environment.

Calcrete aquifers from the Yilgarn region of arid central Western Australia contain an assemblage of obligate groundwater invertebrate species that are each endemic to single aquifers. Fine-scale phylogeographic and population genetic analyses of three sympatric and independently derived species of amphipod (Chiltoniidae) were carried out to determine whether there were common patterns of population genetic structure or evidence for past geographic isolation of populations within a single calcrete aquifer. Genetic diversity in amphipod mitochondrial DNA (cytochrome c oxidase subunit I gene) and allozymes were examined across a 3.5 km(2) region of the Sturt Meadows calcrete, which contains a grid of 115 bore holes (=wells). Stygobiont amphipods were found to have high levels of mitochondrial haplotype diversity coupled with low nucleotide diversity. Mitochondrial phylogeographic structuring was found between haplogroups for one of the chiltoniid species, which also showed population structuring for nuclear markers. Signatures of population expansion in two of the three species, match previous findings for diving beetles at the same site, indicating that the system is dynamic. We propose isolation of populations in refugia within the calcrete, followed by expansion events, as the most likely source of intraspecific genetic diversity, due to changes in water level influencing gene flow across the calcrete.

What lies beneath: molecular phylogenetics and ancestral state reconstruction of the ancient subterranean Australian Parabathynellidae (Syncarida, Crustacea).

The crustacean family Parabathynellidae is an ancient and significant faunal component of subterranean ecosystems. Molecular data were generated in order to examine phylogenetic relationships amongst Australian genera and assess the species diversity of this group within Australia. We also used the resultant phylogenetic framework, in combination with an ancestral state reconstruction (ASR) analysis, to explore the evolution of two key morphological characters (number of segments of the first and second antennae), previously used to define genera, and assess the oligomerization principle (i.e. serial appendage reduction over time), which is commonly invoked in crustacean systematics. The ASR approach also allowed an assessment of whether there has been convergent evolution of appendage numbers during the evolution of Australian parabathynellids. Sequence data from the mtDNA COI and nDNA 18S rRNA genes were obtained from 32 parabathynellid species (100% of described genera and ~25% of described species) from key groundwater regions across Australia. Phylogenetic analyses revealed that species of each known genus, defined by traditional morphological methods, were monophyletic, suggesting that the commonly used generic characters are robust for defining distinct evolutionary lineages. Additionally, ancestral state reconstruction analysis provided evidence for multiple cases of convergent evolution for the two morphological characters evaluated, suggesting that caution needs to be shown when using these characters for elucidating phylogenetic relationships, particularly when there are few morphological characters available for reconstructing relationships. The ancestral state analysis contradicted the conventional view of parabathynellid evolution, which assumes that more simplified taxa (i.e. those with fewer-segmented appendages and setae) are derived and more complex taxa are primitive.

Evidence for population fragmentation within a subterranean aquatic habitat in the Western Australian desert.

The evolution of subterranean animals following multiple colonisation events from the surface has been well documented, but few studies have investigated the potential for species diversification within cavernicolous habitats. Isolated calcrete (carbonate) aquifers in central Western Australia have been shown to contain diverse assemblages of aquatic subterranean invertebrate species (stygofauna) and to offer a unique model system for exploring the mechanisms of speciation in subterranean ecosystems. In this paper, we investigated the hypothesis that microallopatric speciation processes (fragmentation and isolation by distance (IBD)) occur within calcretes using a comparative phylogeographic study of three stygobiontic diving beetle species, one amphipod species and a lineage of isopods. Specimens were sequenced for the mitochondrial cytochrome c oxidase 1 gene from three main sites: Quandong Well, Shady Well (SW) and Mt. Windarra (MW), spanning a 15 km region of the Laverton Downs Calcrete. Phylogenetic and haplotype network analyses revealed that each species possessed a single divergent clade of haplotypes that were present only at the southern MW site, despite the existence of other haplotypes at MW that were shared with SW. IBD between MW and SW was evident, but the common phylogeographic pattern most likely resulted from fragmentation, possibly by a salt lake adjacent to MW. These findings suggest that microallopatric speciation within calcretes may be a significant diversifying force, although the proportion of stygofauna species that may have resulted from in situ speciation in this system remains to be determined.

DNA barcoding of stygofauna uncovers cryptic amphipod diversity in a calcrete aquifer in Western Australia's arid zone.

The arid Yilgarn region of Western Australia contains numerous subterranean calcrete aquifers with unique assemblages of obligate groundwater invertebrates (stygofauna). We aimed to establish a DNA barcoding framework for the macro-invertebrates present in a single calcrete, as a basis for future assessment of biodiversity of the Yilgarn calcretes and for investigating food webs. Intense sampling of a bore field grid in the Sturt Meadows calcrete was undertaken to obtain representatives of the entire macro-invertebrate ecosystem. A 623-bp fragment of the mitochondrial cytochrome c oxidase 1 (COI) gene was used to provide DNA barcodes for stygobiont macro-invertebrates plus terrestrial organisms that are found in the calcrete. Phylogenetic analyses revealed the existence of 12 divergent monophyletic groups of haplotypes. Subterranean amphipods (Chiltoniidae) showed three groups of COI haplotypes with sequence divergences between them of >11%. Allozyme analyses found a large number of fixed allelic differences between these three amphipod groups, indicating that there are three morphologically cryptic species within the Sturt Meadows calcrete. Unlike the sister triplet of dytiscid beetles present, the amphipods are not sister clades and are more closely related to other Yilgarn and non-Yilgarn amphipods than to each other. Our results show that the aquifer contains at least 12 macro-invertebrate species and DNA barcoding provides a useful means for discriminating species in this system.

Fine-scale comparative phylogeography of a sympatric sister species triplet of subterranean diving beetles from a single calcrete aquifer in Western Australia.

Calcrete aquifers in the arid Yilgarn region of central Western Australia are a biodiversity hotspot for stygofauna. A distinct pattern of interspecific size class variation among subterranean dytiscid beetle species has been observed in 29 of these aquifers where either two or three small, medium and/or large sympatric species are found that are in some cases sister species. We used a 3.5 km(2) grid of bores to sample dytiscids on a fine-scale and employed a comparative phylogeographical and population genetic approach to investigate the origins of a sympatric sister species triplet of diving beetles from a single aquifer. Mitochondrial DNA sequence data from the Cytochrome oxidase c subunit I gene revealed that all three species have high levels of haplotype diversity with ancient (approximately 1 million years ago) intra-specific coalescence of haplotypes, but low levels of nucleotide diversity. Population analyses provide evidence for multiple expansion events within each species. There was spatial heterogeneity in the distribution of genetic variation and abundance both within and among the three taxa. Population analyses revealed significant fine-scale differentiation with isolation by distance for Paroster macrosturtensis and P. mesosturtensis, but not the smallest species P. microsturtensis. Haplotype network analyses provided limited or no evidence for past population fragmentation within the large and small species, but substantial historical divergence was observed in P. mesosturtensis that was not spatially structured. A patchy population structure with contemporaneous and historical isolation by distance in the three species is likely to have been a significant isolating and diversifying force, preventing us from ruling out a potential role for allopatric divergence during speciation of this beetle sister triplet.

Cryptic speciation in two widespread subterranean amphipod genera reflects historical drainage patterns in an ancient landscape.

The landscape of the Pilbara region of Western Australia has been relatively unchanged for 100 million years. The ancient river systems of this region might be expected to be sources of isolation and divergence for aquatic species. Hence, the occurrence of widespread groundwater taxa in this landscape offers the opportunity to examine associations between genetic diversity and drainage patterns. Pilbarus and Chydaekata are two widespread genera of subterranean amphipods endemic to the Pilbara, each occupying multiple tributaries. We used molecular data to examine the roles of drainage patterns in structuring genetic diversity. Gene flow within a tributary may be facilitated by the occasional occurrence of these amphipods in springs, which results in their downstream dispersal during episodic flooding. However, tributary boundaries may form hydrological barriers to gene flow, resulting in localised isolation of populations and divergence. Samples of both genera, collected throughout three river basins, were examined for sequence divergence in the cytochrome c oxidase I mitochondrial gene. There was no evidence of contemporary gene flow among populations of either genus, and each tributary contained highly divergent lineages, which were not associated with similar morphological differentiation. This suggests cryptic speciation has occurred, and similar phylogenetic signals in both taxa imply similar evolutionary histories. Surface populations may have been driven into subterranean refugia by the cessation of flow in the rivers, associated with Tertiary climate change, while morphological evolution may have been constrained by stabilising selection. The lack of congruence between molecular diversity and morphology raises important practical issues for conservation.

Production efficiency in small mammal populations.

The data used to analyse the relationship between production (P) and respiration (R) in small mammals (Grodzinski and French 1983) are reanalysed and different conclusions are drawn. Differences in production efficiency cannot be attributed to any particular trophic classe, but in the higher taxonomic categories the murids have higher production effeciency than the rest. However examination of numerous populations of the same species shows that production efficiencies of congeneric species sometimes differ significantly and show differences which exceed those between higher taxa. The same applies to trophic classes. The low slope described for the regression of P on R for Insectivora is due to an aberrant set of data for Sorex araneus. Production efficiency is directly related to litter size in rodents and inversely to survival of adults. This result conflicts with much earlier work and is discussed at length.

The natural history of salmonellae in mammals of the tropical Kimberley region, Western Australia.

Screening for salmonellae was conducted on five occasions within a year on mammals from many different habitats in the Mitchell Plateau area of the tropical Kimberley Division of Western Australia. Some data were obtained from reptiles. Forty-two Salmonella serotypes, two Edwardsiella and one Arizona strain were isolated. Marsupials harbour significantly more (1.6 times) Salmonella serotypes than eutherians. Eleven Salmonella serotypes and one Arizona strain were isolated from reptiles. All Salmonella strains were common to the mammalian species. Varanid lizards showed higher prevalence of salmonellae than other reptilian families. Top carnivores (reptile and marsupial) show relatively high numbers of Salmonella serotypes and prevalence levels. Four mammalian species showed marked seasonal variation in the prevalence of salmonellae and three species in the number of serotypes isolated. Pooled mammalian community data for individual survey sites showed similar seasonal variation which reflected the pattern seen in the predominant host species on the site. Significant seasonal variation in the prevalence of salmonella was concurrent in three of the host species showing maximum prevalence in the wet season (January-summer) followed by a declining prevalence to the end of the dry season (October). Prevalence in the fourth species was also high in January, but continued to increase after the wet season (April) before decreasing as the dry season progressed (July and October). Dendrograms of association showed that the salmonella 'community' does not associate according to any systematic category of the hosts and no evidence of such communities was found using other criteria, such as site or coastal vs inland locations. No evidence was found for host or site specificity of given serotypes. Multiple serotype infections by salmonella were common, with up to three present in mammals and reptiles, and they occurred at random. When hosts in coastal habitats were compared with those in inland sites higher, lower or no difference was found in the prevalence of salmonella, depending upon which host was considered. Sites with similar mammals and vegetation sometimes showed marked differences in salmonella prevalence at a given time. Generally a given Salmonella serotype was lost from the host within congruent to 3 months. Surveys for salmonellae should be designed to include a range of species, replicated vegetation types and, especially, be repeated in several seasons. Edwardsiella tarda was isolated twice from a carnivorous marsupial.

The thermal biology of Geolycosa godeffroyi and other burrow inhabiting Lycosidae (Araneae) in Australia.

Using both field and laboratory data a detailed examination of behavioural thermoregulation is presented for Geolycosa godeffroyi (Koch), a burrow inhabiting Australian wolf spider; comparative data are given for Schizocosa leuckartii (Thorell), Venatrix fuscus (Hogg), Lycosa speciosa Koch, and two undescribed species of Lycosa. Egg sac and spider body temperatures were recorded either by implanted thermocouples or biotelemetry. The burrows permitted the spiders access to a wide range of temperature throughout the day but the spiders maintained temperatures above the upper limit of this range except in the middle of the day in summer. In winter the burrows protected the spiders from extreme cold. G. godeffroyi thermoregulated throughout the year and could achieve temperatures of 38°C when the global radiation exceeded about 260 mW cm-2 d-1. The activity regimes deduced from temperature recording were in accord with those derived using field actographs and photographic recording. The spiders thermoregulated in a manner similar to that used by behaviourally thermoregulating lizards and did so whenever conditions were suitable. They maintained temperatures between 32 and 36°C but the preferred temperature was reduced by lack of food or water, in egg sacs or females carrying egg sacs and in mature males to about 30°C. The data do not support the concept of a refractory zone but suggest two or more set points required for specific physiological or ecological purposes.

The influence of burrowing and thermoregulatory behaviour on the water relations of Geolycosa godeffroyi (Araneae: Lycosidae), an Australian wolf spider.

The water loss from the wolf spider Geolycosa godeffroyi (L. Koch) is presented for a wide range of environmental conditions. Water loss is influenced by the size of the spider, temperature, saturation deficit and by relative humidity per se. The spiders thermoregulatory behaviour more than doubled water loss. Water could not be extracted from near saturated air but was available from the soil when the soil water content was greater than 11%. Due to the spiders burrowing habits both the active and passive use of heat differentials were potential sources of free water.

The food consumption of a wolf spider, Geolycosa godeffroyi (Araneae: Lycosidae), in the Australian Capital Territory.

A method is described to measure the food consumption of a wolf spider, Geolycosa godeffroyi, in the field. The method is based on a calibration curve relating the food consumed in 14 days to the production of components of the excreta, mainly guanine, in the subsequent 7 days. The method was used to measure the food consumption of the spiders in the field over one year. Compared with many arthropods, G. godeffroyi feeds at a high rate but this is probably a reflection of its habit of behavioural thermoregulation.