Cryptic diversity within two widespread diadromous freshwater fishes (Teleostei: Galaxiidae).

Identification of taxonomically cryptic species is essential for the effective conservation of biodiversity. Freshwater-limited organisms tend to be genetically isolated by drainage boundaries, and thus may be expected to show substantial cryptic phylogenetic and taxonomic diversity. By comparison, populations of diadromous taxa, that migrate between freshwater and marine environments, are expected to show less genetic differentiation. Here we test for cryptic diversity in Australasian populations (both diadromous and non-diadromous) of two widespread Southern Hemisphere fish species, Galaxias brevipinnis and Galaxias maculatus. Both mtDNA and nuclear markers reveal putative cryptic species within these taxa. The substantial diversity detected within G. brevipinnis may be explained by its strong climbing ability which allows it to form isolated inland populations. In island populations, G. brevipinnis similarly show deeper genetic divergence than those of G. maculatus, which may be explained by the greater abundance of G. maculatus larvae in the sea allowing more ongoing dispersal. Our study highlights that even widespread, 'high-dispersal' species can harbour substantial cryptic diversity and therefore warrant increased taxonomic and conservation attention.

Cherax latimanus sp. nov., a new burrow-dwelling freshwater crayfish (Decapoda, Parastacidae) from the mid-Murray River catchment, Australia.

A new species of freshwater crayfish in the southern hemisphere family Parastacidae is described from the Murray Darling Basin (MDB), eastern Australia. The Swamp Yabby, Cherax latimanus sp. nov., is found in the mid-Murray River area of the states of New South Wales and Victoria and is only the third species of Cherax found naturally occurring in the MDB. It is morphologically distinguished from all other species of Cherax by the presence of marginal mesial dactylar basal spines and extensive ventral propodal setation. It is found in sympatry with the widespread but morphologically similar Common Yabby, Cherax destructor Clark, 1936, but can be distinguished by a suite of morphometric and meristic characters that include a broad rostrum lacking an apical spine, very broad chelae with deep punctations on the lateral propodal edge and ventral propodal setation, two prominent dactylar teeth, one mesial carpal spine, dorsal meral spines present, no setae on carpal mesial margin, cervical spines absent or tiny, and by genetic data. Cherax latimanus is further diagnosed by its distinctive biology: it is not found in permanent aquatic habitats such as streams or billabongs, spending the majority of its time in extensive, terrestrial, burrow networks containing some water, in ephemerally wet habitats such as drainage lines, roadside drains, depressions, swamps and cleared areas of pasture in lowland to foothill areas. It is occasionally found in stream habitats but only during large flood events when burrow systems are inundated.

Multispecies models reveal that eDNA metabarcoding is more sensitive than backpack electrofishing for conducting fish surveys in freshwater streams.

Environmental DNA (eDNA) sampling can provide accurate, cost-effective, landscape-level data on species distributions. Previous studies have compared the sensitivity of eDNA sampling to traditional sampling methods for single species, but similar comparative studies on multispecies eDNA metabarcoding are rare. Using hierarchical site occupancy detection models, we examined whether key choices associated with eDNA metabarcoding (primer selection, low-abundance read filtering and the number of positive water samples used to classify a species as present at a site) affect the sensitivity of metabarcoding, relative to backpack electrofishing for fish in freshwater streams. Under all scenarios (teleostei and vertebrate primers; 0%, 0.1% and 1% read filtering thresholds; one or two positive samples required to classify species as present), we found that eDNA metabarcoding is, on average, more sensitive than electrofishing. Combining vertebrate and teleostei markers resulted in higher detection probabilities relative to the use of either marker in isolation. Increasing the threshold used to filter low-abundance reads decreased species detection probabilities but did not change our overall finding that eDNA metabarcoding was more sensitive than electrofishing. Using a threshold of two positive water samples (out of five) to classify a species as present typically had negligible effects on detection probabilities compared to using one positive water sample. Our findings demonstrate that eDNA metabarcoding is generally more sensitive than electrofishing for conducting fish surveys in freshwater streams, and that this outcome is not sensitive to methodological decisions associated with metabarcoding.

Revision of the genus Hannia (Teleostei, Terapontidae), with description of a new species, Hannia wintoni, from the Kimberley, Western Australia.

The systematics of the genus Hannia Vari 1978, endemic to freshwater habitats of remote north-western Australia, is revised in light of recent collections in the region and a molecular study of the group that identified an undescribed candidate species. A new freshwater fish species (Hannia wintoni sp. nov) is described based on analysis of multiple nuclear genetic markers (53 allozyme loci), mitochondrial DNA sequence data (601 bp cytochrome b) and morphology (examination of a suite of 66 morphometric and meristic characters). Head profile, postorbital length, maximum length, preopercular spines and pectoral-fin rays are characters that best distinguish H. wintoni sp. nov from its only congener, H. greewayi. While the existing description of H. greenwayi is robust and accurate, we present a number of additional characters that enhance to the original description, based on type and fresh material. Information on the known distribution, habitats and conservation status of the two species is summarised. The new species is a narrow-range endemic.

Revision of the Australian Wet Tropics endemic rainbowfish genus Cairnsichthys (Atheriniformes: Melanotaeniidae), with description of a new species.

The freshwater melanotaeniid genus Cairnsichthys is endemic to a relatively small area of specialised habitat within the Wet Tropics bioregion of north-eastern Queensland, Australia. It was previously considered as monotypic, including only a single species, C. rhombosomoides (Nichols Raven, 1928). The recent discovery of an apparently-isolated population in the Daintree rainforest, approximately 120 km north of the known range extent, prompted a detailed investigation of its taxonomic status using a combined lines of evidence approach. We provide compelling evidence from multiple nuclear genetic markers (52 allozyme loci), mitochondrial DNA sequence data (1141 bp cytochrome b) and morphology (examination of a suite of 38 morphometric and meristic characters) that supports north-south splitting of C. rhombosomoides. Accordingly, we describe the northern population as a distinct species, C. bitaeniatus sp. nov., on the basis of 25 specimens, 34.7-65.6 mm SL. The new species differs morphologically primarily by having a more slender and narrow shape, featuring a flatter, straighter predorsal profile and shorter second dorsal fin base; possession of slightly smaller scales, reflected in higher counts of lateral scales and predorsal scales; typically more vertebrae; and colour differences including a more robust, short black stripe across the upper operculum, a pronounced yellow patch on the anteroventral body and usually a more conspicuous second dark stripe on the lower body, with adult males generally having yellowish compared to reddish fins. We also provide a generic diagnosis for Cairnsichthys and a redescription of C. rhombosomoides. Information on the known distribution, habitats and conservation status of species in the genus is summarised, the new species being of particular concern as a narrow range endemic with specific environmental requirements.

Genome-wide SNPs resolve a key conflict between sequence and allozyme data to confirm another threatened candidate species of river blackfishes (Teleostei: Percichthyidae: Gadopsis).

Conflicting results from different molecular datasets have long confounded our ability to characterise species boundaries. Here we use genome-wide SNP data and an expanded allozyme dataset to resolve conflicting systematic hypotheses on an enigmatic group of fishes (Gadopsis, river blackfishes, Percichthyidae) restricted to southeastern Australia. Previous work based on three sets of molecular markers: mtDNA, nuclear intron DNA and 51 allozyme loci was unable to clearly resolve the status of a putative fifth candidate species (SWV) within Gadopsis marmoratus. Resolving the taxonomic status of candidate species SWV is particularly critical as based on IUCN criteria this taxon would be considered Critically Endangered. After all filtering steps we retained a subset of 10,862 putatively unlinked SNP loci for population genetic and phylogenomic analyses. Analyses of SNP loci based on maximum likelihood, fastSTRUCTURE and DAPC were all consistent with the previous and updated allozyme results supporting the validity of the candidate Gadopsis species SWV. Immediate conservation actions should focus on preventing take by anglers, protection of water resources to sustain perennial reaches and drought refuge pools, and aquatic and riparian habitat protection and improvement. In addition, a formal morphological taxonomic review of the genus Gadopsis is urgently required.

A review of Galaxiella pusilla (Mack) (Teleostei: Galaxiidae) in south-eastern Australia with a description of a new species.

The dwarf galaxias, Galaxiella pusilla (Mack), is a small, threatened freshwater fish from coastal south-eastern Australia. Recent genetic studies, using multiple nuclear and mitochondrial DNA markers, found substantial differences between populations in western Victoria and south Australia ('west region') compared to eastern Victoria, Flinders Island, and Tasmania ('east region') that suggest the presence of a cryptic species. Morphological measurements and meristic counts from multiple populations within each region were undertaken to investigate potential differences between regions. Several characters, found to discriminate between individuals in the regions and to be diagnostic for two taxa, were used to describe a new species, Galaxiella toourtkoourt, for the west region. This is only the second species in the Galaxiidae to exhibit sexual dimorphism. The original description of Galaxiella pusilla, based on five specimens, is revised following examination of a large number of individuals. Both species are considered nationally threatened and are categorised as 'endangered'; the revised distribution of G. pusilla s.s. is reduced by approximately 60%. A number of inconsistencies in the most recent revision of the genus Galaxiella are also corrected.

Fifteen from one: a revision of the Galaxias olidus Günther, 1866 complex (Teleostei, Galaxiidae) in south-eastern Australia recognises three previously described taxa and describes 12 new species.

The systematics of the Galaxias olidus hyper-species complex from freshwater habitats in south-eastern, mainland Australia is revised. Galaxias olidus Günther 1866 is redescribed, Galaxias fuscus Mack 1936 and Galaxias ornatus Castelnau 1873, previously synonymised with G. olidus (sensu lato), are reinstated as valid taxa and redescribed, and 12 taxa are described as new: Galaxias aequipinnis sp. nov., Galaxias arcanus sp. nov., Galaxias brevissimus sp. nov., Galaxias gunaikurnai sp. nov., Galaxias lanceolatus sp. nov., Galaxias longifundus sp. nov., Galaxias mcdowalli sp. nov., Galaxias mungadhan sp. nov., Galaxias oliros sp. nov., Galaxias supremus sp. nov., Galaxias tantangara sp. nov., and Galaxias terenasus sp. nov. These species are morphologically similar and, whilst there is extensive overlap in meristic counts and morphometric characters, each can be diagnosed by unique combinations of characters, including allozyme loci and colour pattern; morphological diagnosis is improved greatly if based on freshly formalin-fixed material. Galaxias schomburgkii Peters 1868, Galaxias bongbong Macleay 1881, Galaxias kayi Ramsay & Ogilby 1886 and Galaxias oconnori Ogilby 1912 are retained as junior synonyms of G. olidus (sensu stricto). The types for Galaxias findlayi Macleay 1882 are lost and no specimens matching its description were collected or examined from the Mt. Kosciuszko region; it is also currently retained as a junior synonym of Galaxias olidus s.s. The species G. terenasus sp. nov. and G. arcanus sp. nov. are the most morphologically specialised in the complex and G. olidus s.s remains the most morphologically variable species. It also remains the most widespread taxon, though its previously known distribution is reduced, particularly in the south-west of its range. Nine species are narrow-range endemics, known from one, or only a few, locations, and these restricted distributions most probably reflect the fragmentation and reduction of former ranges caused by the effects of alien salmonids. Eleven species are of conservation concern, most are considered critically endangered.

Global biodiversity assessment and hyper-cryptic species complexes: more than one species of elephant in the room?

Several recent estimates of global biodiversity have concluded that the total number of species on Earth lies near the lower end of the wide range touted in previous decades. However, none of these recent estimates formally explore the real "elephant in the room", namely, what proportion of species are taxonomically invisible to conventional assessments, and thus, as undiagnosed cryptic species, remain uncountable until revealed by multi-gene molecular assessments. Here we explore the significance and extent of so-called "hyper-cryptic" species complexes, using the Australian freshwater fish Galaxias olidus as a proxy for any organism whose taxonomy ought to be largely finalized when compared to those in little-studied or morphologically undifferentiated groups. Our comprehensive allozyme (838 fish for 54 putative loci), mtDNA (557 fish for 605 bp of cytb), and morphological (1963-3389 vouchers for 17-58 characters) assessment of this species across its broad geographic range revealed a 1500% increase in species-level biodiversity, and suggested that additional taxa may remain undiscovered. Importantly, while all 15 candidate species were morphologically diagnosable a posteriori from one another, single-gene DNA barcoding proved largely unsuccessful as an a priori method for species identification. These results lead us to draw two strong inferences of relevance to estimates of global biodiversity. First, hyper-cryptic complexes are likely to be common in many organismal groups. Second, no assessment of species numbers can be considered "best practice" in the molecular age unless it explicitly includes estimates of the extent of cryptic and hyper-cryptic biodiversity. [Galaxiidae; global estimates; hyper-diverse; mountain galaxias; species counts; species richness.].